Pressure regulated continuously variable volume container for fluid delivery.
16 claims: 16 independent, 0 dependent
- 1CLAIMS REIVINDICACIONES IM IM 7Γ 7Γ R x x msTL?pn R xx msTL? Pn INDUSTRIAL ---- A flow cytometer comprising:INDUSTRIAL ----Un citómetro de flujo que comprende: a variable volume container that has a flexible wall;un contenedor de volumen variable que tiene una pared flexible;b. an amount of fluid established within said variable volume container;b. una cantidad de fluido establecida dentro de dicho contenedor de volumen variable;c. a gas inlet element that allows to provide a quantity of gas that exerts a pressure in a range of 0.706 kg / cm2 (10 psi) to 14.1kg / cm2 (200 psi) on the outer surface of said flexible wall of said variable volume container to generate a stream of fluid in a conduit fluidically coupled between said variable volume container and a flow path of said flow cytometer;c. un elemento de entrada de gas que permite proporcionar una cantidad de gas que ejerce una presión en un intervalo de 0.706 kg/cm2 (10 psi) a 14.1kg/cm2 (200 psi) sobre la superficie exterior de dicha pared flexible de dicho contenedor de volumen variable para generar una corriente de fluido en una conducto fluídícamente acoplado entre dicho contenedor de volumen variable y una trayectoria de flujo de dicho citómetro de flujo;d. a particle source that intermittently transports a particle in said fluid stream;d. una fuente de partículas que transporta de manera intermitente una partícula en dicha corriente de fluido;and. at least one lighting source that generates at least one incident light beam in said particle contained in said fluid stream for a duration of time;e. al menos una fuente de iluminación que genera al menos un haz de luz incidente en dicha partícula contenida en dicha corriente de fluido por una duración de tiempo;F. an oscillator acting in said fluid stream to generate a plurality of drops in said fluid stream, wherein at least one of said plurality of drops contains said particle;f. un oscilador que actúa en dicha corriente de fluido para generar una pluralidad de gotas en dicha corriente de fluido, en donde al menos una de dicha pluralidad de gotas contiene dicha partícula;-57IMPI @ 53. -57IMPI @53. NíSTrTUTO MEXICANO tVj— ' ! '2 ηΕΙΑΕΕΟΗΕΟΑΙ' i,. . , .INDUSTRIAL _ níSTrTUTO MEXICANO tVj—' ! '2 ηΕΙΑΕΕΟΗΕΟΑΙ' i , . . , , .INDUSTRIAL _ g. al menos un detector Adaptado para recibir una emisión generada por dicha partícula contéΓΪIüaen”dicha gota que varía en base a al menos una característica de partícula. g. at least one detector Adapted to receive an emission generated by said particle conté gotaIüaen ”said drop that varies based on at least one particle characteristic.
- 2The flow cytometer as described in claim 1, wherein said variable volume container comprises a tubular body having a first end and a second end each hermetically formed. 2. El citómetro de flujo como se describe en la reivindicación 1, en donde dicho contenedor de volumen variable comprende un cuerpo tubular que tiene un primer extremo y un segundo extremo cada uno formado herméticamente.
- 3The flow cytometer as described in claim 1, wherein said fluid is selected from the group consisting of a flowable fluid in the flow path of said flow cytometer, a water, a buffer, a coating fluid, a fluid of flow cytometer coating, a liquid reagent, a cleaning solution, and a solvent. 3. El citómetro de flujo como se describe en la reivindicación 1, en donde dicho fluido se selecciona del grupo que consiste de un fluido fluible en la trayectoria de flujo de dicho citómetro de flujo, un agua, un amortiguador, un fluido de revestimiento, un fluido de revestimiento de citómetro de flujo, un reactivo líquido, una solución de limpieza y un solvente.
- 4El citómetro de flujo como se describe en la reivindicación 1, en donde dicha cantidad de gas se selecciona del grupo que consiste de un gas atmosférico, una mezcla de gases, una mezcla de gases que tienen presiones parciales seleccionadas, un gas purificado, un gas de nitrógeno, un gas helio. Four. The flow cytometer as described in claim 1, wherein said amount of gas is selected from the group consisting of an atmospheric gas, a mixture of gases, a mixture of gases having selected partial pressures, a purified gas, a gas nitrogen, a helium gas.
- 5Claim 1, group consisting of sperm cell flow cytometer as described in wherein said particle is selected from that of a cell, a sperm cell, a labeled, a stained sperm cell, a sperm cell having a fluorochrome to a 5. El reivindicación 1, grupo que consiste célula espermática citómetro de flujo como se describe en la en donde dicha partícula se selecciona del de una célula, una célula espermáticas, una etiquetada, una célula espermática teñida, una célula espermática que tiene un enlace de fluorocromo a un INSTITUTO MEXICANO DE LA PROPIEDAD c de 'Β«έί MEXICAN INSTITUTE OF PROPERTY c de 'Β «έί ADN nuclear, cromosoma, un fragmento de proteína. Nuclear DNA, chromosome, a fragment of protein. a component obtained from nucleic acid, a protein, UTV AUN;ΟΤΓ "7ΏξϊΦ7 ~ τΠΤ ' un componente obtenido a partí ácido nucleico, una proteína, UTV AUN;ΟΤΓ“7ΏξϊΦ7~τΠΤ' ADN, un fragmento de ARN y un fragmento de DNA, a fragment of RNA and a fragment of
- 6The flow cytometer as described in claim 1, further comprising a receptacle in which said variable volume container having said flexible wall has a location. 6. El citómetro de flujo como se describe en la reivindicación 1, que comprende además un receptáculo en el cual dicho contenedor de volumen variable que tiene dicha pared flexible tiene una ubicación.
- 7The flow cytometer as described in claim 6, wherein said amount of gas exerting pressure on said flexible wall of said variable volume container is collected between an inner surface of said receptacle and said outer surface of said flexible wall of said variable volume container. 7. El citómetro de flujo como se describe en la reivindicación 6, en donde dicha cantidad de gas que ejerce una presión sobre dicha pared flexible de dicho contenedor de volumen variable se recolecta entre una superficie interior de dicho receptáculo y dicha superficie exterior de dicha pared flexible de dicho contenedor de volumen variable.
- 8The flow cytometer as described in claim 7, wherein said receptacle comprises a material selected from the group consisting of stainless steel, aluminum, plastic, cardboard and cardboard. 8. El citómetro de flujo como se describe en la reivindicación 7, en donde dicho receptáculo comprende un material seleccionado del grupo que consiste de un acero inoxidable, un aluminio, un plástico, un cartón y una cartulina.
- 9The flow cytometer as described in claim 8, wherein said receptacle comprises a coating fluid tank for said flow cytometer. 9. El citómetro de flujo como se describe en la reivindicación 8, en donde dicho receptáculo comprende un tanque de fluido de revestimiento para dicho citómetro de flujo.
- 10The flow cytometer as described in claim 1, further comprising a nozzle responsive to said oscillator acting with said fluid stream to 10. El citómetro de flujo como se describe en la reivindicación 1, que comprende además una tobera sensible a dicho oscilador que actúa con dicha corriente de fluido para -59IMPI -59IMPI INSTITUTO MEXICANO -|R MEXICAN INSTITUTE - | R DE LA MiOfIBDAD CSto-^áaí >li J INDUSTRIAL generar dicha pluralidad de gotas. OF THE CSO- ^ áaí> li J INDUSTRIAL MiOfIBILITY generate said plurality of drops.
- 11El citómetro de flujo como aesb'rTB^^STT'-la reivindicación 1, en donde al menos un haz de luz incidente en dicha partícula contenido en dicha gota por una duración de tiempo comprende al menos una haz de láser. eleven. The flow cytometer as aesb'rTB ^^ STT'-claim 1, wherein at least one beam of light incident on said particle contained in said drop for a duration of time comprises at least one laser beam.
- 12The flow cytometer as described in claim 11, wherein said at least one laser beam comprises a pulsed laser beam. 12. El citómetro de flujo como se describe en la reivindicación 11, en donde dicho al menos un haz de láser comprende un haz de láser pulsado.
- 13The flow cytometer as described in claim 1, wherein said emission generated by said particle that varies based on at least one particle characteristic comprises a fluorescent emission generated by said particle. 13. El citómetro de flujo como se describe en la reivindicación 1, en donde dicha emisión generada por dicha partícula que varía en base a al menos una característica de partícula comprende una emisión fluorescente generada por dicha partícula.
- 14The flow cytometer as described in claim 13, wherein said at least one particle characteristic comprises an amount of DNA contained in said particle. 14. El citómetro de flujo como se describe en la reivindicación 13, en donde dicha al menos una característica de partícula comprende una cantidad de ADN contenido en dicha partícula. dicha partícula comprende una cantidad de ADN contenido en un said particle comprises a quantity of DNA contained in a -60 sperm bearing the Y chromosome. -60espermatozoide que lleva el cromosoma Y. c c 17. The flow cytometer as claimed in claim 1, wherein said at least one single detector. 17. El citómetro de flujo como reivindicación 1, en donde dicho al menos un un solo detector. IMP IMP INSTITUTO MEXICANO «IA MOEISBAD IMCtSTFIAt se describe en la detector comprende MEXICAN INSTITUTE «IA MOEISBAD IMCtSTFIAt is described in the detector comprises 18. The flow cytometer as described in claim 1, wherein said at least one detector comprises two detectors. 18. El citómetro de flujo como se describe en la reivindicación 1, en donde dicho al menos un detector comprende dos detectores. 19. The flow cytometer as described in claim 18, wherein said at least two detectors comprise a first photomultiplier tube and a second photomultiplier tube. 19. El citómetro de flujo como se describe en la reivindicación 18, en donde dichos al menos dos detectores comprenden un primer tubo fotomultiplicador y un segundo tubo fotomultiplicador. 20. El citómetro de flujo como se describe en la reivindicación 1, que comprende además un generador de señal acoplado a dicho al menos un detector diferencialmente sensible de cada dicha emisión generada por cada dicha partícula. twenty. The flow cytometer as described in claim 1, further comprising a signal generator coupled to said at least one differentially sensitive detector of each said emission generated by each said particle. particle comprises a quantity of DNA contained in said particle. partícula comprende una cantidad de ADN contenida en dicha partícula. 2. 3. The cytometer! flow as described in 23. El citómetro! de flujo como se describe en la -61 I Ni PI -61 I Ni PI ERsrrrvTo mexicano PE LA tROHEDAD reivindicación 21, que comprende además un cT&^fficaWe^-í^e partícula que aísla cada dicha partícula eh'babe a id—prggerteira- de dicha al menos una característica de partícula en un contenedor de recolección. Mexican ERsrrrvTo PE LA tROHEDAD claim 21, further comprising a cT & ^ fficaWe ^ -í ^ e particle which isolates each said particle eh'babe a id-prggerteira- from said at least one particle characteristic in a collection container. 24. The flow cytometer as described in claim 23, wherein said particle classifier separates sperm cells carrying the X chromosome from those carrying the Y chromosome. 24. El citómetro de flujo como se describe en la reivindicación 23, en donde dicho clasificador de partícula separa las células espermáticas que llevan el cromosoma X de las que llevan el cromosoma Y. 25. The flow cytometer as described in claim 1, wherein said flexible wall comprises at least two layers, wherein a first layer has a surface compatible with said amount of gas and a second layer has a surface compatible with said liquid. 25. El citómetro de flujo como se describe en la reivindicación 1, en donde dicha pared flexible comprende al menos dos capas, en donde una primera capa tiene una superficie compatible con dicha cantidad de gas y una segunda capa tiene una superficie compatible con dicho líquido. 26. The flow cytometer as described in claim 25, wherein said first layer comprises a material selected from the group consisting of a polypropylene, a polyethylene, a fluorocarbon, a styrene and a polycarbonate. 26. El citómetro de flujo como se describe en la reivindicación 25, en donde dicha primera capa comprende un material seleccionado del grupo que consiste de un polipropileno, un polietileno, un fluorocarburo, un estireno y un policarbonato. 27. The flow cytometer as described in claim 25, wherein said second layer comprises a material selected from the group consisting of a polypropylene, a polyethylene, a fluorocarbon, a styrene and a polycarbonate. 27. El citómetro de flujo como se describe en la reivindicación 25, en donde dicha segunda capa comprende un material seleccionado del grupo que consiste de un polipropileno, un polietileno, un fluorocarburo, un estireno y un policarbonato. 28. The flow cytometer as described in 28. El citómetro de flujo como se describe en la -62IMPIí ^ c rmrmrro **FORMER* ¿^ G .. -62IMPIí^c rmrmrro **EX*¿^g .. Claim 25, wherein said cantidagl of EfaaND ^ wxe reivindicación 25, en donde dicha cantidagl de EfaaND^wxe ►. ·· * pressure on said outer surface Η p_ H -i z- hn p-,T. said variable volume container is collected between said first layer and said second layer. ►.·· * presión sobre dicha superficie exterior Η p_ H -i z- h n p-,T. de dicho contenedor de volumen variable se recolecta entre dicha primera capa y dicha segunda capa. 29. The flow cytometer as described in claim 25, wherein said first layer and said second layer are joined. 29. El citómetro de flujo como se describe en la reivindicación 25, en donde se unen dicha primera capa y dicha segunda capa. 30. The flow cytometer as described in claim 25, wherein said flexible wall further comprises at least one intermediate layer. 30. El citómetro de flujo como se describe en la reivindicación 25, en donde dicha pared flexible comprende además al menos una capa intermedia. 31. The flow cytometer as described in claim 30, wherein said at least one intermediate layer comprises a material selected from the group consisting of a polypropylene, a polyethylene, a fluorocarbon, a styrene and a polycarbonate. 31. El citómetro de flujo como se describe en la reivindicación 30, en donde dicha al menos una capa intermedia comprende un material seleccionado del grupo que consiste de un polipropileno, un polietileno, un fluorocarburo, un estireno y un policarbonato. 32. A flow cytometry method, comprising the steps of:32. Un método de citometría de flujo, que comprende las etapas de: a. proporcionar un contenedor de volumen variable que tiene una pared flexible;to. providing a variable volume container that has a flexible wall;b. establecer una cantidad de fluido en dicho contenedor de volumen variable;b. setting a quantity of fluid in said variable volume container;c. ejercer una cantidad de presión con una cantidad de gas en un intervalo de 0.706 kg/cm2 (10 psi) a 14.1kg/cm2 (200 psi) sobre una superficie exterior de dicha c. exert an amount of pressure with an amount of gas in an interval of 0.706 kg / cm2 (10 psi) to 14.1kg / cm2 (200 psi) on an outer surface of said -63 IMPI -63 IMPI INSTTTUTC MEXICANO 'Dt LA ^ tOMEDAD flexible wall of said container of vol | te (in variaBTe;INSTTTUTC MEXICANO ‘ Dt LA^tOMEDAD pared flexible de dicho contenedor de voli|te(en variaBTe;5 flow;5 flujo;and. intermittently transporting a plurality of particles in said fluid stream from a particle source;e. transportar de manera intermitente una pluralidad de partículas en dicha corriente de fluido desde una fuente de partículas;F. generate oscillations in said current of f. generar oscilaciones en dicha corriente de 10 fluid containing said plurality of particles to establish a plurality of drops;10 fluido que contiene dicha pluralidad de partículas para establecer una pluralidad de gotas;g. establecer una de dicha pluralidad de partículas en una de dicha pluralidad de gotas;g. establishing one of said plurality of particles in one of said plurality of drops;h. illuminate said particle in each of h. iluminar dicha partícula en cada una de
- 1515 dicha pluralidad de gotas por una duración de tiempo;e fifteen said plurality of drops for a duration of time;and i. detectar una emisión proveniente de dicha partícula que varía en base a al menos una característica de partícula. i. detecting an emission from said particle that varies based on at least one particle characteristic. 33. The flow cytometry method as 33. El método de citometría de flujo como se
- 1620 describe en la reivindicación 32, en donde dicha etapa de proporcionar un contenedor de volumen variable comprende la etapa de proporcionar un cuerpo tubular flexible que tiene un extremo superior y un extremo inferior cada uno formado herméticamente. twenty discloses in claim 32, wherein said step of providing a variable volume container comprises the step of providing a flexible tubular body having an upper end and a lower end each hermetically formed. -6434. The method of claim 32, in generating a fluid stream in a fluidically coupled conduit between said variable volume container and a flow path of a flow cytometer comprises generating a fluid stream from the group consisting of a flowable fluid in the flow path of said flow cytometer, a water, a buffer, a coating fluid, a coating fluid of the flow cytometer, a liquid reagent, cleaning solutions and a solvent. -6434. El método de describe en la reivindicación 32, en generar una corriente de fluido en un conducto fluídicamente acoplado entre dicho contenedor de volumen variable y una trayectoria de flujo de un citómetro de flujo comprende generar una corriente de fluido del grupo que consiste de un fluido fluíble en la trayectoria de flujo de dicho citómetro de flujo, un agua, un amortiguador, un fluido de revestimiento, un fluido de revestimiento del citómetro de flujo, un reactivo líquido, unas soluciones de limpieza y un solvente. 35. The flow cytometry method as described in claim 32, wherein said step of exerting an amount of pressure with an amount of gas on said flexible wall of said variable volume container further comprises selecting said amount of gas from the group consisting of of an atmospheric gas, a mixture of gases, a mixture of gases having selected partial pressures, a purified gas, a nitrogen gas, a helium gas. 35. El método de citometría de flujo como se describe en la reivindicación 32, en donde dicha etapa de ejercer una cantidad de presión con una cantidad de gas sobre dicha pared flexible de dicho contenedor de volumen variable comprende además seleccionar dicha cantidad de gas del grupo que consiste de un gas atmosférico, una mezcla de gases, una mezcla de gases que tienen presiones parciales seleccionadas, un gas purificado, un gas de nitrógeno, un gas helio. 36. The flow cytometry method as described in claim 32, wherein said step of intermittently transporting a particle in said fluid stream from a particle source comprises intermittently transporting a particle in said fluid stream selected from the group consisting of a 36. El método de citometría de flujo como se describe en la reivindicación 32, en donde dicha etapa de transportar de manera intermitente una partícula en dicha corriente de fluido a partir de una fuente de partículas comprende transportar intermitentemente una partícula en dicha corriente de fluido seleccionada del grupo que consiste de una -65 IMPIég ^ cell, a sperm cell, a labeled ceiuaiwsTMAfesp ^ nrraTica, a sperm cell He has. or.1:.. ι ^ ιιιρπιτΒτκτΐ.— obtained from a cell, a chromosome, a nucleic acid, a protein, a DNA, an RNA, a fragment of DNA, a fragment of RNA, and a fragment of protein. -65 IMPIég^ célula, una célula espermaticas, una ceiuaiwsTMAfesp^nrraTica etiquetada, una célula espermática Ha. u.1:.. ι^ιιιρπιτΒτκτΐ.— obtenido de una célula, un cromosoma, un ácido nucleico, una proteína, un ADN, un ARN, un fragmento de ADN, un fragmento de ARN y un fragmento de proteína. 37. The flow cytometry method as described in claim 32, further comprising the step of providing a receptacle in which said variable volume container has a location. 37. El método de citometría de flujo como se describe en la reivindicación 32, que comprende además la etapa de proporcionar un receptáculo en el cual dicho contenedor de volumen variable tiene una ubicación. 38. The flow cytometry method as described in claim 37, wherein said step of exerting an amount of pressure with an amount of gas on said flexible wall of said variable volume container further comprises the step of collecting said amount of gas between an inner surface of said receptacle and said outer surface of said variable volume container. 38. El método de citometría de flujo como se describe en la reivindicación 37, en donde dicha etapa de ejercer una cantidad de presión con una cantidad de gas sobre dicha pared flexible de dicho contenedor de volumen variable comprende además la etapa de recolectar dicha cantidad de gas entre una superficie interior de dicho receptáculo y dicha superficie exterior de dicho contenedor de volumen variable. 39. The flow cytometry method as described in claim 32, wherein said stage illuminating said particle in each of said plurality of drops for a duration of time comprises the stage of illuminating said 39. El método de citometría de flujo como se describe en la reivindicación 32, en donde dicha etapa que ilumina dicha partícula en cada una de dicha pluralidad de gotas por una duración de tiempo comprende la etapa de iluminar dicha -66ΙΜΡΙ -66ΙΜΡΙ INSTITUTO MEXICANO MEXICAN INSTITUTE INSTITUTO MEXICANO detect an emission from said particle qqlL vSwS ^ Mfn based on at least one characteristic of pajXí ^ uÍA ™ e © H ^ -ende ---- le · stage of receiving a fluorescent emission that varies based on a difference in amount of DNA inside the cell. INSTITUTO MEXICANO detectar una emisión a partir de dicha partícuíawiqttaL vSwS^Mfn base a al menos una característica de pajXí^uÍA™e©H^-ende----le· etapa de recibir una emisión fluorescente que varía en base a una diferencia en cantidad de ADN dentro de la célula. 5 41. The flow cytometry method as described in claim 40, further comprising the step of analyzing said particle based on said emission that varies based on said at least one particle characteristic. 5 41. El método de citometría de flujo como se describe en la reivindicación 40, que comprende además la etapa de analizar dicha partícula en base a dicha emisión que varía en base a dicha al menos una característica de partícula. 42. The flow cytometry method as described in claim 41, further comprising the step of separating said particle based on the presence of said at least one particle characteristic. 42. El método de citometría de flujo como se 10 describe en la reivindicación 41, que comprende además la etapa de separar dicha partícula en base a la presencia de dicha al menos una característica de partícula. 43. The flow cytometry method as described in claim 42, wherein said step of 43. El método de citometría de flujo como se describe en la reivindicación 42, en donde dicha etapa de 15 separar dicha partícula en base a la presencia de dicha al menos una característica de partícula comprende separar las células espermáticas que llevan el cromosoma X de las que llevan el cromosoma Y. fifteen separating said particle based on the presence of said at least one particle characteristic comprises separating the sperm cells that carry the X chromosome from those that carry the Y chromosome. 44. The flow cytometry method as 44. El método de citometría de flujo como se 20 describe en la reivindicación 32, que comprende además la etapa de proporcionar dicha pared flexible con al menos dos capas. twenty describes in claim 32, further comprising the step of providing said flexible wall with at least two layers. 45. El método de citometría de flujo como se describe en la reivindicación 44, que comprende además la etapa de proporcionar una primera capa compatible con dicha cantidad Four. Five. The flow cytometry method as described in claim 44, further comprising the step of providing a first layer compatible with said amount -67 de gas y proporcionar una segunda fluido. -67 gas and provide a second fluid. cap capa IMPI IMPI INSTITUTO MEXICANO comf£H¡*g^ MEXICAN INSTITUTE comf £ H¡ * g ^ 46. The flow cytometry method as described in claim 45, further comprising the step 46. El método de citometría de flujo como se describe en la reivindicación 45, que comprende además la etapa 5 of collecting said amount of gas between said first layer and said second layer. 5 de recolectar dicha cantidad de gas entre dicha primera capa y dicha segunda capa. 47. The flow cytometry method as described in claim 45, further comprising the step of attaching said first layer to said second layer. 47. El método de citometría de flujo como se describe en la reivindicación 45, que comprende además la etapa de unir dicha primera capa a dicha segunda capa. 10 48. The flow cytometry method as described in any of claims 45, 46, or 47, wherein said first layer comprises a material selected from the group consisting of polypropylene, polyethylene, fluorocarbon, styrene, and polycarbonate . 10 48. El método de citometría de flujo como se describe en cualquiera de las reivindicaciones 45, 46, ó 47, en donde dicha primera capa comprende un material seleccionado del grupo que consiste de polipropileno, un polietileno, un fluorocarburo, un estireno y un policarbonato. 15 49. El método de citometría de flujo como se describe en cualquiera de las reivindicaciones 45, 46, 47 ó 48, en donde dicha segunda capa comprende un material seleccionado del grupo que consiste de polipropileno, un polietileno, un fluorocarburo, un estireno y un policarbonato. fifteen 49. The flow cytometry method as described in any one of claims 45, 46, 47 or 48, wherein said second layer comprises a material selected from the group consisting of polypropylene, a polyethylene, a fluorocarbon, a styrene and a polycarbonate. 20 50. El método de citometría de flujo como se describe en la reivindicación 45, que comprende además proporcionar 5al menos una capa de material intermedio entre dicha primera capa y dicha segunda capa. twenty 50. The flow cytometry method as described in claim 45, further comprising providing 5at least one layer of intermediate material between said first layer and said second layer.
Independent claims16
269 paragraphs in 92 sections, as filed
(54) Title: CONSTANTLY VARIABLE VOLUME CONTAINER REGULATED BY PRESSURE FOR THE SUPPLY OF FLUID.
(54) Title: PRESSURE REGULATED CONTINUOUSLY VARIABLE VOLUME CONTAINER FOR FLUID DELIVERY.
(57) Summary
The present invention relates to a flow cytometer comprising: a. a variable volume container that has a flexible wall; b. an amount of fluid established within said variable volume container; c. a gas inlet element that allows to provide an amount of gas that exerts a pressure in a range of 0.706 kg / cm2 (10 psi) to 14.1kg / cm2 (200 psi) on the outer surface of said flexible wall of said container of variable volume to generate a fluid stream in a fluidically coupled conduit between said variable volume container and a flow path of said flow cytometer; d. a particle source that intermittently transports a particle in said fluid stream; and. at least one lighting source that generates at least one incident light beam in said particle contained in said fluid stream for a duration of time; F. an oscillator acting in said fluid stream to generate a plurality of drops in said fluid stream, wherein at least one of said plurality of drops contains said particle; g. at least one detector adapted to receive an emission generated by said particle contained in said drop that varies based on at least one particle characteristic.
(57) Abstract
A fluid handling and delivery system useful in generating a fluid stream (7) in the flow path (8) of microfluidic device (16).
I KNOW
Institute
Mexican Property
Industrial
<img file="MX339262B_D0001.tif" />
PATENT TITLE NO. 339262
Owner (s): XY, LLC; CHATA BIOSYSTEMS, INC.
Address: 22575 State Highway 6 South, Navasota, Texas, 77868, USA
Name: CONSTANTLY VARIABLE VOLUME CONTAINER REGULATED BY PRESSURE FOR THE SUPPLY OF FLUID
Classification:
Inventor (s):
lnt.CI.8: B01L3 / 00; F16K17 / 40; G01N35 / 08
EDWIN DEAN NEAS; JERALD EDWARD KUIKEN; JOHN LOUIS SCHENK; THOMAS BOYD GILLIGAN
<img file="MX339262B_D0002.tif" />
Wildebeest
MX / f / 2
».......................... IF> 1 * 1»
Country:
US
International filing date:
December 2005 to Patent Number: 293376
PRIORITY
Date: TT Number:
December 2004
11/004,382
Security: Twenty years • Vencí cha reference patent pursuant to article 23 of the Law of the<sup>1 </sup>ntada from the ugly one and the presentation of nachos to ¡in 2025 | s articles 1, 2 fraction V, 6 fraction III, and S9 of the Industrial Property Law. industrial, the present patent has a validity of twenty years. Non-expendable, it will be subject to the payment of the fee to keep in force the number: 2 di itorga con fundadan »
<img file="MX339262B_D0003.tif" />
Who subscribes to the present Industrial Opportunity (Daily title lo! ice with Official Fund of the Federation i n'o en J
01/01/2004, 06/16/2005, 2 01/2006, 03/05/2009, 06/01/01 201 a), subsection iii) 4 «and 12th fractions I and lll of Regulation T / 07/2004, formed on 07/01/2002, of the Organic Statute Jiel Institu 3 ° and
07/08/2004 and 09/07/2007); art
Mexican Industrial Property and
by ios a-tioulos 6 * fractions lll and 7 ° bis 2 of i prmada e. 08/02/1994, 10/25/1996, 12/26/1997, 08/28/2--0 27/012012 and 09®4 / 2'.12); Articles 1, Industrial Jledad (DOFJ Siso a), sub clause iii) 161
7712/1999, amended on 10/10/2002,
Law of
05/07/1999, section V 12/4/1999, jnes I and lll and> 07/09/2004, General
Deputies, Coordinator, Divisional Directors, Head of the Regional Offices, Divisional Deputy Directors, Departmental Coordinators and other subordinates of the Mexican Institute of Industrial Property. (DOF 12/15/1999, amended on 02/04/2000, 07/29/2004, 08/04/2004 and 09/13/2007).
<img file="MX339262B_D0004.tif" />
Issue Date: May 18, 2016
DIVISIONAL TOR FOR EXAMINATION OF PATENT FUND CTRICA AREAS AND REGISTRIES OF INDUSTRIAL DESIGNS AND MODELS
PEDR
<img file="MX339262B_D0005.tif" />
ArenaiNo. 550, Floor 1,
Coi. Santa María Tepepan town. Xcchimilco. CP 16020.
Mexico City
Tel (55) 53 34 Oí 00 www iinni mx
<img file="MX339262B_D0006.tif" />
<img file="MX339262B_D0007.tif" />
MX / 2016/38422
<img file="MX339262B_D0008.tif" />
CONSTANTLY VARIABLE VOLUME CONTAINER REGULATED BY
PRESSURE FOR THE SUPPLY OF FLUID
This Patent Application of the International Patent Cooperation Treaty claims the benefit of the
United States Non-Provisional Patent Application No. 11 / 004,382, filed on December 3, 2004, incorporated herein by reference.
I. TECHNICAL FIELD
A continuously variable pressure regulated volume container for fluid handling and supply. Specifically, a pressure regulated variable volume container useful for generating a stream of fluid in the flow path of various types of microfluidic devices such as flow cytometers or liquid chromatographs.
II. BACKGROUND
Flow cytometry, liquid chromatography, and other microfluidic devices are prominent tools used in basic and applied research and in commercial manufacturing processes. These microfluidic systems are routinely used to analyze, separate, isolate or purify biological particles, such as cells, organelles, chromosomes, deoxyribonucleic acids (DNA), ribonucleic acids (RNA), DNA fragments, RNA fragments, proteins, fragments of protein.
IMPI
MEXICAN INSTITUTE OF PROPERTY
INDUSTRIAL
<img file="MX339262B_D0009.tif" />
peptides, oligonucleotides, or the like. -—---- '
Specifically with respect to applications in flow cytometry or the use of flow separation devices, biological particles, such as cells (which can be modified with one or a plurality of types or classes of ligands, labels or fluorescent inks) or particles carriers (which may contain particles such as antibodies or oligonucleotides or the like), they can be analyzed and separated to isolate individual cells or biological particles or subpopulations of cells or biological particles, which have one or a plurality of common characteristics. As the field of flow cytometry has matured, an increased emphasis has been placed on the retention of the biological function (s) of isolated biological cells or particles.
Flow cytometers can also be used to analyze and separate a mixture of non-biological particles. For example, non-biological particles can be differentially modified with analyte-specific reagents and reacted with a heterogeneous mixture of biological particles or analytes. The non-biological particles loaded with the biological particles or specific analytes of the corresponding reagent can then be differentiated and isolated with the flow separation system. Flow separation applications of this type
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY can provide sequence analysis dtí splCógiéS <sup>l</sup>S ”for genes similar to those of a multi-gene microarray analysis using a flat surface such as a microscope slide, to present different analyte-specific reagents such as antibodies, oligonucleotides, aptamers or the like, for one or more biological particles of a heterogeneous biological mixture.
To maintain the biological function (s) of living cells during analysis, separation, purification or collection, cells are transported in fluids prepared to have certain characteristics related to purity, pH, ion concentration, osmolality , buffer capacity, availability of nutrients and the like. With respect to certain applications, these fluids must be prepared with validated water to be free of foreign agents, pyrogens or the like; or with chemicals obtained from suppliers validated in accordance with regulatory specifications such as cGMP guidelines, 510K guidelines, ISO-9000 guidelines, batch registration documentation, documentation of the main drug file or the like.
Specifically with respect to chromatographic systems, the fluids used to transport and separate biological particles are often mixtures
<img file="MX339262B_D0010.tif" />
IMPI
MEXICAN INSTITUTE <sup>W</sup> OF THE ΡΚΟΡ1ΕΟΑΠ ° INDUSTRIAL
<img file="MX339262B_D0011.tif" />
5 · purified from solvents and solutes in water. You can opt 1 ΐ - the variable mix between two or more fluids to establish differential gradients of salt concentration, pH, solvent ratios or the like to selectively release the particles from a variety of solid substrates to effect particle separation biological in subpopulations based on one or more characteristics of the particle.
The feature of chromatographic systems is the relatively large volume of fluid used to separate mixtures of different particle (s) or population (s) of particles into individual particles or purified subpopulations of particles that are then isolated in a relatively small volume of fluid. Typically, many liters of an elution buffer can be collected in a plurality of individual fractions each containing a few milliliters of the desired isolated product in one or some of such fractions. Fluid preparation and handling to support chromatographic applications must be carried out reliably by appropriately trained technicians. Any inaccuracy in the preparation of such fluids can lead to a significant loss of the operating time of the chromatograph or the total or part loss of the unpurified mixed particle (s) or the (s) population (s) of particles or the
TO
IMPI
MEXICAN INSTITUTE OF PROPERTY
INDUSTRIAL
<img file="MX339262B_D0012.tif" />
purified individual particle (s) <p —1 a (s) - .e ^ b ·· population (s) of the particles of interest.
Understandably, extensive research has been conducted resulting in numerous and varied types of microfluidic devices, fluids used with such microfluidic devices, and methods of making and using such microfluidic devices to separate biological and non-biological particles as described above, or from another way. However, significant problems remain unresolved with respect to establishing and maintaining consistency in the preparation, handling and supply of fluids to and in the ducts of such microfluidic devices.
A significant problem with the conventional supply of fluids to microfluidic devices may be contamination of the fluid. Transfer of the fluid from a fluid container to a microfluidic device and additional transfer of the fluid through various analytical passages may require the generation of hydrostatic pressure. Typically, a pump supplies the hydrostatic pressure required to move a fluid into and out of the lines of a microfluidic device.
Positive displacement pumps, for example, absorb fluid from one side of the pump body and
IMPI
MEXICAN INSTITUTE DEIAPROPIEQAD industrial
<img file="MX339262B_D0013.tif" />
using valves, pistons, rotors, vanes or the like. „Force the fluid to the other side of the pump. In this process, the fluid can come into contact with the internal surfaces of the pump, depositing non-biological or biological materials, microbial or other infectious agents, which can remain inside the body of the pump. In this way, the surfaces of the pump body can become a source of contamination for the subsequent volume of fluid transferred through the pump body.
Peristaltic pumps apply pressure to the outer surface of a conformable conduit to act on the fluids contained within the conformable conduit. Peristalsis of the conformable conduit transfers fluid in one direction within the body of the conformable conduit. An advantage of the peristaltic pump may be that the fluids do not contact the surfaces of the peristaltic pump. However, peristaltic pumps have disadvantages in that they cannot form very high pressures, they can tend to create oscillating variations in hydrostatic pressure, they can be expensive to build and maintain, and recurrent peristalsis of the formable conduit can cause progressive deformation or degradation of the material of the duct that can leak, drain or leak into the fluid.
<img file="MX339262B_D0014.tif" />
IMPI
Another significant conventional problem of fluids to microfluidic devices may be the use of a gas or gas mixtures, such as air, argon, nitrogen, helium or the like, to pressurize the upper space of a fluid container to initiate and maintain a flow. of fluid in the conduits of the microfluidic device. The use of pressurized gas (es) or atmospheric gas pressure in contact with the fluid in the container can result in the formation of bubbles in the fluid paths of the device. Since microfluidic devices have small diameter flow paths and the biological particles carried in the fluid stream are also small in size, even very small or fine bubbles formed in the flow path can affect the volume and laminar flow of the Fluid within the flow paths can cause failure of certain types of pumps and can result in analytical errors. Even bubbles invisible to the naked eye can be problematic with regard to the proper performance of a microfluidic device.
One mechanism by which unwanted bubbles can spontaneously form in the flow path of a microfluidic device may be a change in the concentration of the dissolved gas in the liquid stream followed by the formation of bubbles. For example, a
MEXICAN INSTITUTE AND PROPERTY
INDUSTRIAL coating fluid container puttener 'an amount of coating fluid to operate a flow cytometer for a long duration of time, sometimes more than 72 hours. With a discharge pressure of more than four atmospheres or in certain applications in more than 6 atmospheres, the dissolved nitrogen content of the fluid can increase dramatically as the gases in the liquid move toward equilibrium with the gases in the upper space of the container.
Subsequently, when the gas pressure in the liquid is reduced, bubbles can form. The reduction in gas pressure may come from operator inspection or manipulation of the amount of fluid remaining in the coating fluid container. Alternatively, as the fluid flows through the conduits of the microfluidic device, the fluid pressure can become substantially lower to equalize the operating pressure of the microfluidic flow path. Under these conditions bubbles can form and travel within the flow path of the microfluidic device. Alternatively, the surface tension of the bubble may allow it to adhere to the surfaces of the analytical components of the microfluidic device. The attached bubbles can also serve as condensation nuclei where bubbles fuse
<img file="MX339262B_D0015.tif" />
additional small or where bubbles can enter ^ -ai -.— additional dissolved gas.
The position of such bubbles that divide between a surface adherent phase and the suspended phase of fluid is determined by the size of the bubble and the rate of fluid flow at that point in the apparatus. Microfluidic devices, flow cells, and flow cytometers commonly have regions in the flow path where the flow is not laminar, where the flow rate is low, and where bubbles tend to form. For example, microfluidic devices may have filters that purposely restrict fluid flow to facilitate removal of unwanted particles or aggregates. Bubbles are often collected on the upstream side of such filters, effectively reducing the filter surface area available for the fluid. Also, because gas can easily move through a filter, either as dissolved gas or as bubbles that may be smaller than the filter exclusion dimension, bubbles can also accumulate on the opposite side of the filter.
Unwanted bubbles can also be formed in a microfluidic device by direct transfer of the pressurized gas into the flow path of the microfluidic device.
For example, when
IMPI
Mexican Institute of Industrial Property
<img file="MX339262B_D0016.tif" />
Coating fluid containers of conventional flow cytometry run out of fluid or when the amount of fluid is low and the container is not level or when the coating fluid container is pumped, tilted or agitated, pressurized gas can directly enter to the flow path of the device. When pressurized gas enters the flow path of a microfluidic device directly, the bubbles may be larger and in certain circumstances may interrupt the flow of the fluid completely, alter the flow characteristics, or remain located in the flow path of the microfluidic device . If the microfluidic device or flow path is not disposable, it may take a significant amount of time to dislodge or discharge the unwanted bubbles from the flow path.
Another problem related to the use of pressurized gas in contact with liquids to generate a fluid stream in microfluidic devices may be an increased concentration of oxygen in solution. For example, live sperm cells in the presence of media containing energy sources may exhibit a metabolic rate limited by the content of dissolved oxygen. During and after separation of the flow of sperm cells it may be advantageous to have a rate
IMPI
MEXICAN INSTITUTE INDUSTRIAL PROPERTY
High can be generated
<img file="MX339262B_D0017.tif" />
Viable but low metabolic rates, dissolved oxygen concentrations by balancing the coating fluid with pressurized oxygen-containing gases, and their use can result in disadvantageously high metabolic rates in sperm cells during flow analysis or flow separation processes.
A similar problem with using atmospheric gases or pressurized gases in contact with fluids to generate a fluid stream can be increased amounts of water introduced into anhydride solvents or other water sensitive fluids used within microfluidic devices.
Another similar problem with using atmospheric gases or pressurized gases in contact with fluids to generate a fluid stream may be the reaction of certain gases with the fluid or the particles carried in the fluid.
Another significant problem with conventional fluid preparation for use with microfluidic devices or chromatography systems may be that the quality of the available water or the quality of the chemical solvent may be unacceptably low to make standard solutions for certain applications. Although there are numerous and varied methods to increase the quality of
<img file="MX339262B_D0018.tif" />
<img file="MX339262B_D0019.tif" />
water, the cost of use can be unacceptably high the water source contains a certain level of one or a plurality of materials, substances or pathogens. This problem can be compounded by the use of specialized fluids for applications in basic research, cell-based clinical therapy, or pharmaceutical production that may require higher quality fluids with regard to formulation accuracy, batch-to-batch consistency, and material-free , particles, inorganic and organic substances, pathogens, chemicals or the like unwanted contaminants. Particularly with respect to fluids that buffer or provide carbon sources to maintain cellular function, high-quality water may be essential to prevent or reduce pathogen growth to acceptably low levels.
Several of these problems are identified by Neas US Patent No. 6,729,369, which is directed to preparing large volumes of specialized sterile fluids in a single geographic location where high quality water and chemicals are available. Fenced flexible containers are then used to transport the prepared sterile specialized fluids to the location where the fluids are used. However, Neas et al. Does not address the problem of establishing a pressurized fluid stream in the flow path of any
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY
<img file="MX339262B_D0020.tif" />
microfluidic device such as a flu-cytometer. liquid chromatograph or the like.
Another significant problem with the conventional supply of fluids to microfluidic devices may be cleaning, removal of unused amounts of fluid, and sterilization of fluid containers. Flow cytometers can consume between approximately 200 milliliters to approximately 800 milliliters of coating fluid per hour and typically operate between approximately one hour and twenty-four hours for a single procedure. Coating fluid tanks or containers typically contain between about five to about ten liters of coating fluid and if a procedure is interrupted or terminated, it is often inconvenient to store the unused coating fluid in the coating fluid container for use. in the same procedure at a later date, Because the coating fluid tank may be needed for other procedures or the coating fluid may undergo microflora growth or microfine still if stored. Even if the coating fluid is stored, it can often be kept between 4-10 ° C and must then be rebalanced at warm temperatures before further use.
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY
Many products are distributed in the broad consumer market as fluid containers that are opened for use and accordingly, the fluids in the container begin to interact with the atmosphere. With respect to certain fluids, interaction with the atmosphere can be harmful to the stability or consistency of the fluid. For example, paint or other surface coating products can start to cure when exposed to the atmosphere by moving toward equilibrium with the volume of the atmosphere in the container. As such, a portion of unused paint in a container can form a thin layer of film. Another example may be free radical mediated rancification of edible oils such as olive oil, polyunsaturated vegetable oils or the like, accelerated by molecular oxygen.
Many fluids are dispensed into small pressurized containers that supply the fluid through an orifice that causes the fluid to disperse when it leaves the container. Common examples are spray paint cans, hair sprayers, deodorants, insecticides, pesticides, herbicides, or the like. A disadvantage of small pressurized containers is that there is a limited number of acceptable propellants that are both inert to the reaction with the fluid (s).
<img file="MX339262B_D0021.tif" />
IMPI ^
MEXICAN INSTITUTE
OF PROPERTY Vj »—M
INDUSTRIAL content (s) as yet benign to the environment. ———
For larger-scale application, these fluids are typically contained in reusable containers that can be pressurized with a hand pump or with air compressors. In addition to the problems discussed above regarding gas interaction with fluids, there are additional disadvantages related to the safety of cleaning large containers of the remaining fluids and the disposal of the remaining fluids.
The current invention provides fluid delivery devices and fluid delivery methods that address each of the aforementioned problems with conventional technology in the specific area of microfluidic devices as well as the broader consumer market.
III. DESCRIPTION OF THE INVENTION
Accordingly, a broad objective of the invention may be to eliminate exposure of the fluid (s) that are supplied in the flow path of a microfluidic device or chromatography systems to external sources of contamination. One aspect of this objective of the invention may be to isolate fluids that are supplied to the flow path of a microfluidic device from movement toward equilibrium with atmospheric gases, gas mixtures, or gas partial pressures either at atmospheric pressure or at pressures. greater than
IMPI
MEXICAN INDUSTRIAL PROPERTY UTILITY
<img file="MX339262B_D0022.tif" />
atmospheric. A second aspect of the objective eotc of the invention may be to isolate fluids that are supplied to the flow path of a microfluidic device from exposure to non-biological materials or surfaces, such as pump surfaces, dust, cleaning compositions, or the similar; or to biological substances or surfaces that can introduce pathogens, bacteria, viruses, spores, cells, proteins, nucleic acids, tissues, blood, semen, urine, faeces or the like. A third aspect of this objective of the invention may be to maintain a sterile fluid to be supplied to the flow path of a microfluidic device.
Another broad object of the invention may be to provide a container that has a continuously adjustable volume with respect to the amount of fluid contained within so that a stream of fluid can be supplied to a microfluidic device. One aspect of this objective of the invention may be to provide a container that has a continuously variable volume in response to pressure exerted on the outer surface which enables the inner surface of the container to act on the fluid contained within to generate a fluid stream in an exit. A second aspect of this objective of the invention may be to provide a flexible wall capable of withstanding a gas pressure of between approximately
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL MONEDAD
<img file="MX339262B_D0023.tif" />
4.92 Kg / cm<sup>2</sup> (70 pounds per square inch (psi)) v 7-03. Kg / cm<sup>2</sup> (100 psi) to generate a fluid stream in the flow path of a microfluidic device of approximately 1.76 Kg / cm<sup>2</sup> (25 psi) up to approximately 3.52 Kg / cm<sup>2</sup> (50 psi). Naturally, for certain applications the amount of pressure exerted on the flexible wall may be greater and for certain applications the amount of pressure may be less. A third aspect of this objective of the invention may be to supply from a container having continuously variable volume in response to the pressure exerted by a quantity of gas or liquid a current of fluid towards the flow path of a microfluidic device, such as a flow cytometer or liquid chromatography, in which particles can be transported for analysis, separation, purification or otherwise manipulated as desired.
Another broad object of the invention may be to provide an improvement in or re-adaptation to conventional fluid container technology which further includes a container having a continuously adjustable volume with respect to the amount of fluid contained within, such that a fluid stream it can be supplied to a microfluidic device. One aspect of this objective of the invention may be to retrofit the coating fluid tanks of the conventional flow cytometer to
The MEXICAN INDUSTRIAL PROPERTY WSTITVTO will further include a container that has a continuously adjustable volume with respect to the amount of fluid contained within so that a stream of fluid can be supplied to a flow cytometer. A second aspect of this objective of the invention with respect to liquid chromatographs may be to retrofit conventional liquid phase containers to further include a container that has a continuously adjustable volume with respect to the amount of fluid contained therein, so that the Fluid stream can be supplied directly to the separation column or to the high pressure pump of the liquid chromatograph.
Another broad aspect of the invention may be to establish or maintain a desired concentration of a gas or gases dissolved in the supplied fluid in the flow path of a microfluidic device such that the particles (either biological or non-biological) are exposed to the concentration or level of gas (is) necessary (s) or desired); or exposure of particles to certain gases, gas mixtures or gas partial pressures, increased water content or the like may be avoided.
Another broad aspect of the invention may be to provide fluids that are prepared to conform to the specifications of a particular microfluidic device or
IMPI
MEXICAN INSTITUTE OF INOUSTWAL PROPERTY
<img file="MX339262B_D0024.tif" />
method of using the microfluidic device and transferring it to a container having a continuously variable volume according to the invention. Such containers prepared at a first geographic location can then be shipped to various other geographic locations to maintain the consistency of the fluids used by the microfluidic devices at each location.
Another broad object of the invention may be to provide a receptacle of substantially fixed configuration in which a quantity of gas or liquid can be supplied to act on the surface of a container having a variable volume to supply fluid to the flow path of a microfluidic device. . One aspect of this broad embodiment of the invention may be to provide a receptacle of substantially fixed configuration having a plurality of compartments allowing a plurality of fluids to be simultaneously supplied to one or more microfluidic devices or containers.
Another broad embodiment of the invention may be to provide a flow cytometer device or chromatographic system and methods for using such a flow cytometer device or chromatographic system that uses fluids separated from the surfaces of the coating fluid tank and the gases supplied to the tank. of coating fluid.
Mexican Institute of Industrial Property
Another broad objective of the invention "may be to provide fluids and methods for supplying fluids to the flow paths of microfluidic devices that are compatible with the isolation or purification of cells or other particles or substances for reintroduction into a human or animal. There are a significant number of issues that arise regarding the prevention of transmission of infection or disease when cells, particles, or substances are isolated by microfluidic devices. Infectious particles or other agents can range in size from prions that can be a few tenths of a nanometer, to virus particles that can be hundreds of nanometers, to yeasts, fungi, molds and bacteria that can be several hundred nanometers to many micrometers in size. Once a sample of cells, particles, or other substances are contaminated with such infectious particles, it can be very difficult to remove them. In some cases, agents such as preservatives or antibiotics are acceptable, but in most products that are used in animals and humans, government regulations require the use of production methods that can be validated to produce biological cells, particles, substances or chemicals free of all such particles or foreign infectious agents. The current invention facilitates preparation, shipping.
<img file="MX339262B_D0025.tif" />
<sup>21</sup> IMPI
MEXICAN INSTITUTE OF PROPERTY m · INDUSTRIAL
P »Storage, handling and use of validated sterile solutions free of particles or foreign agents that can be supplied under pressure to the flow cytometer, flow cell or other microfluidic devices or chromatographic systems to generate cells, particles or other substances free of infectious agents or unwanted others.
Specific examples of such treatments or therapies may be the isolation of specific hematopoietic stem cells from the bone marrow with the procedure of separating abnormal or cancerous cells from normal cells and the reinsertion of non-cancerous or normal cells back into the bone marrow. ; isolating certain white blood cells or cancer blood cells and modifying such cells with certain conjugates and adjuvants that allow the cells to re-insert (dead or alive) as a form of therapeutic vaccination; the isolation of very rare cells, such as fetal cells, from blood, such as maternal blood, containing a very small number of such fetal cells, for the purpose of carrying out genetic analysis such as the chain reaction of polymerase (PCR), determining the genotype or halotype of such fetal cells, with a minimal genetic background from much more abundant genetic content of maternal blood cells; cell isolation
IMPI
MEXICAN INSTITUTE ^ ELA mOTlEDAD
INDUSTRIAL
<img file="MX339262B_D0026.tif" />
such as sperm cells from vaginal fluids for the purposes of analyzing the genetic makeup of sperm cells; flow separation of mammalian sperm cells to generate enriched populations that have the X chromosome and that have the viable sperm Y chromosome or flow separation of enriched sperm cells for certain genetic characteristics for further use in assisted reproductive techniques such such as in vitro fertilization, intra-cytoplasmic sperm injection, artificial insemination or the like.
Another broad object of the invention may be to provide a container having a continuously variable volume with respect to a quantity of conformable material contained therein, such that such a comfortable material such as; water, a fluid for a microfluidic device; a coating fluid for flow cytometry; A food; a drink; a food ingredient; a drink ingredient; a liquid detergent; a liquid pesticide or herbicide; a pharmaceutical solvent such as rubbing alcohol; u a toiletries product such as a shampoo, body wash, hair spray or hair gel; or the like can be handled, supplied, circulated in the flow path of a conduit or otherwise used with only the desired contact with the
<img file="MX339262B_D0027.tif" />
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY
Btfi.
atmosphere or other partial pressures '' of gases and without being released into the atmosphere or other partial pressures of gases, unless desired. One aspect of this modality is the provision of a large variable volume of containers containing specialized concentrates that are useful in the processing industry that formulates and produces fluidic products for consumers and may be beneficial from new methods for accurate delivery. and clean of fluid products, at controlled quantities, in the products of which they compose.
Naturally, additional objects of the invention are described through other areas of the specification, drawings and claims.
IV. BRIEF DESCRIPTION OF THE DRAWINGS
Figure IA shows an embodiment of the invention that provides a variable pressure regulated volume container that supplies a stream of fluid in response to a quantity of gas acting on the outer surface.
Figure IB shows a cross section through the flexible wall of one embodiment of the pressure regulated variable volume container.
Figure 1C shows a cross section through the flexible wall of the alternative embodiment of the pressure regulated variable volume container.
Figure ID shows a cross section through
<img file="MX339262B_D0028.tif" />
through regulated alternative.
IMPI
MEXICAN INSTITUTE • OF PROPERTY, 'INDUSTRIAL the flexible wall of the second modality of the container of variable pressure volume
Figure 1E shows a cross section through the flexible wall of a third alternative embodiment of the pressure regulated variable volume container.
Figure 2A shows a conventional coating fluid tank for supplying the coating fluid (s) to a flow cytometer.
Figure 2B shows an embodiment of the invention for supplying the coating fluid (s) to a flow cytometer in which a conventional coating fluid tank is retrofitted to receive an amount of gas that acts on the outer surface of a container of variable volume.
Figure 2C shows an alternative embodiment of the invention for supplying the coating fluid (s) to a flow cytometer in which a conventional coating fluid tank is retrofitted to receive an acting amount of gas the outer surface of a container of variable volume.
Figure 3A shows an embodiment of the invention in which a receptacle of substantially fixed configuration receives an amount of gas acting on the surface.
IMPI
MEXICAN INSTITUTE OF PROPERTY
INDUSTRIAL
<img file="MX339262B_D0029.tif" />
exterior of a variable volume container.
Figure 3B shows an alternative embodiment of the invention in which a receptacle of substantially fixed configuration receives an amount of gas that acts on the outer surface of a container of variable volume.
Figure 4A shows an embodiment of the invention in which a plurality of receptacles each receives an amount of. gas that acts on the outer surface of a container of variable volume to generate a plurality of fluid streams.
Figure 4B shows an alternative embodiment of the invention, in which a plurality of receptacles each receives a quantity of gas that acts on the outer surface of a container of variable volume to generate a plurality of fluid streams.
Figure 5 shows an embodiment of the flow cytometer of the invention in which a stream of fluid can be generated, in the flow path of the flow cytometer, from a container of variable volume acting by a quantity of gas.
Figure 6 shows a bivariate graph of sperm cells transported in a fluid stream generated according to the invention, differentiated into populations that have the X chromosome and that have the
-26- IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY chromosome Y. _____________
Figure 7 shows an embodiment of the invention in which a plurality of containers of variable volume each containing a quantity of fluid are configured on a sheet of columns and rows.
Figure 8 illustrates a plurality of flow paths operable with the embodiment of the invention shown by Figure 7.
V. MODE (S) FOR CARRYING OUT THE INVENTION
Generally an amount of fluid is located within a variable volume container that has a flexible wall that acts with the amount of fluid in response to the gas pressure exerted on the outer surface to generate a fluid stream in the flow path of a conduit.
Now referring primarily to Figure 1, one embodiment of the invention can provide a variable volume container (1) having a flexible wall (2) that acts on a quantity of fluid (3) within the variable volume container (1) in response to a quantity of pressure (4) exerted on the outer surface (5) of the flexible wall (2) by a quantity of gas (6). The amount of pressure (4) exerted on the outer surface (5) of the flexible wall (2) continuously adjusts the volume of the variable volume container
<img file="MX339262B_D0030.tif" />
<img file="MX339262B_D0031.tif" />
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY (1) to act on the quantity of fluid (3) to generate a stream of fluid (7) (either continuous flow or discontinuous flow) in the flow path of a duct (8) . As in certain embodiments of the invention, the variable volume container (1) can partly be of a substantially rigid configuration and partly a flexible wall (2). That portion of the variable volume container (1) that provides the flexible wall (2) can act on the amount of fluid (3) within the variable volume container (1) in response to the amount of pressure (4) exerted on the outer surface (5) of the flexible wall by the amount of gas (6) to generate a fluid stream (7).
The fluid (3) within the variable volume container (1) broadly encompasses, without limitation, any fluid, liquid, composition, mixture, phase, product or other fluidizable material in the flow path of the conduit (8) by continuously adjusting the volume of the variable volume container (1) in response to the amount of pressure (4) exerted on the outer surface (5) of the flexible wall (2). The numerous and varied fluidisable fluids in the duct flow path (8) (the duct flow path includes numerous and varied configurations corresponding to the wide range of applications for the invention and without limitation include the
<img file="MX339262B_D0032.tif" />
<img file="MX339262B_D0033.tif" />
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY miciuf luídieir 'qtre flow paths or conduits typically have an internal diameter of approximately one millimeter or less) includes without limitation: water, a solvent, a solution, a buffer, a liquid chromatography solution, a fluid in which biological particles can be transported, a fluid in which non-biological particles can be transported, a fluid in which cells can analyzed, a fluid in which sperm cells can be analyzed, a fluid in which sperm cells can be separated into populations carrying the Y chromosome and carrying the X chromosome, a flow cytometric coating fluid, a flow cytometric coating fluid in which non-biological particles can be transported, a flow cytometric coating fluid in which biological particles can be transported, a cytometric coating fluid flow in which cells are transported, a flow cytometric coating fluid in which sperm can be transported, a flow cytometric coating fluid in which stained sperm, paint, pesticides, pastes, adhesives, organic solvents, pesticides, food products, beverages, and various permutations and combinations thereof can be transported.
Now referring mainly to Figure IB,
<img file="MX339262B_D0034.tif" />
the variable volume container (1) can provide a flexible wall (2) on which the ac:
exerts an amount of pressure (4). The flexible wall (2) can comprise a layer of material (9) that has enough flexibility to adjust the volume of the variable volume container (1) in response to the amount of pressure (4) exerted by the amount of gas (6) on the outer surface (5). The layer of material (9) can be selected to provide an interior surface (10) compatible with the fluid 1 (3) contained within the variable volume container (1) and provide an exterior surface (5) compatible with the amount of gas ( 6) that exerts the amount of pressure (4) on it. With respect to certain embodiments of the invention, the material layer can be further selected to avoid or minimize the transfer of leachable or transferable materials from the material layer (9) to the fluid (3) contained by the variable volume container (1 ). The material layer (9) can also be selected to avoid or minimize the transfer of the amount of gas (6) through the material layer (9) into the fluid contained by the variable volume container (1). Without limiting the numerous and varied materials that can be used in accordance with the invention, preferred embodiments of the invention can utilize a layer of material (9) such as polypropylene, a
- 30 - IMPI
MEXICAN INSTITUTE
OF THE PROPERTY
INDUSTRIAL i, i polyethylene, a nylon, a fluorine or carbide,. a neotiiciiiu, polycarbonate, a metal foil, a laminated paper, a biodegradable polymer, a wax paper, or bonded layers thereof in various permutations and combinations. The material layer 9 may include a material coating, such as an oxygen barrier, a water barrier, alternate layers of a surface-filling polymer, and a ceramic (eg Barix) or the like.
Now referring mainly to Figure 1C, as in other embodiments of the invention, the flexible wall (2) can comprise two layers of material. The first layer (11) establishes the outer surface (5) compatible with the amount of gas (6) that exerts a pressure (4) on the flexible wall (2) and a second layer (12) that provides an inner surface (10) ) compatible with the fluid (3) inside the variable volume container (1). The first layer 11 can be selected from materials such as a polypropylene, a polyethylene, a fluorocarbon, a styrene, a polycarbonate, a Mylar® film, an oxygen barrier, a water barrier or the like. The second layer (12) can be selected from the same or a different material than the first layer (11) such as a polypropylene, a polyethylene, a fluorocarbon, a styrene, a polycarbonate, a water barrier or an oxygen barrier ( for example Barix) or the like. Either or both of the
<img file="MX339262B_D0035.tif" />
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY
<img file="MX339262B_D0036.tif" />
first layer (11) or the second layer (12) can-JjaelwÁ * - «« temás · a reinforcing element (48) such as individual fibers, threads, strands, a mesh, net or the like that can be made of a material reinforcement such as nylon, cotton, carbon fiber, metal strand, plastic strand, or the like.
As in certain embodiments of the invention, the first layer (11) and the second layer (12) of a flexible wall (2) can be slidably clutched, while in like other embodiments of the invention the first layer (11) and the second layer (12) can be fixedly clutched. The fixed clutch between the first layer (11) and the second layer (12) can be generated by the use of an adhesive layer (13) or another type of layer or other process that induces a surface of the first layer (11) and a surface of the second layer (12) to adhere to each other.
Now referring mainly to Figure ID, as in other particular embodiments of the invention, a gas collection element (14) can be interposed between the first layer (11) and the second layer (12). As in these embodiments of the invention, the amount of gas (6) that exerts a quantity of pressure (4) on the outer surface (5) of the variable volume container (1) is collected in the gas collection element (14 ) and exerts a quantity of pressure (4) on the second layer (12) that
......- 1 --- 32 IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY acts on the liquid (3) contained within to generate the
<img file="MX339262B_D0037.tif" />
fluid stream (7). The first layer (11) acts to adjust the volume or pressure (or both) of the amount of gas (6) within the gas collection element (14) to the necessary or desired amount. As in these embodiments of the invention, in which the first layer (11) does not have to function as part of a flexible wall (2) of the variable volume container (1), the first layer (11) can have a substantially configuration fixed formed from a material such as a plastic, a fiberglass, a glass, a metal, a steel, a polycarbonate, an acrylic, a polypropylene, a vinyl, a fluorinecarbon, a carbon fiber or the like.
Now referring primarily to Figure 1E, other embodiments of the invention may further include a flexible wall (2) having at least one intermediate layer (15) located between the first layer (11) and the second layer (12). The at least one intermediate layer (15) can be selected from a material such as a polypropylene, a polyethylene, a fluorocarbon, a styrene, a polycarbonate, a Mylar® film, a ceramic layer, an oxygen barrier (or other gas ), a water barrier or the like. Additional embodiments of the invention may further provide the gas collection element (14) (similar to Figure ID) interposed between the first layer
IMPI
INSTITUTO MEXICANO DE LA RROÍIEDAD INDUSTRIAL (11) and the at least one intermediate layer (15) or as for others
<img file="MX339262B_D0038.tif" />
particular embodiments of the invention the gas collection element (14) (similar to Figure ID) can be interposed between the second layer (12) and the at least one intermediate layer (15). Where the gas collection element (14) is interposed between the first layer (11) and the at least one intermediate layer (15), the first layer (11) can be of substantially fixed configuration as described above. In those embodiments of the invention, in which the gas collection element (14) is interposed between the second layer (12) and the at least one intermediate layer (15), any of the first layer (11) or the al minus an intermediate layer (15) or both, can have a substantially fixed configuration, while the second layer (12) provides enough flexibility to allow the variable volume container (1) to continuously adjust the volume in response to the amount of pressure (4) exerted by the amount of gas (6) on the flexible wall ( 2). As in those embodiments of the invention, in which the liquid (3) clutches the inner surface (10) of the second layer (12) and the quantity of gas (6) exerts a quantity of pressure (4) on the outer surface (5) of the first layer (11), then the first layer (11), the intermediate layer (15) and the second layer (12) can have enough flexibility to allow the variable adjustment volume of the
IMPI
MEXICAN INSTITUTE OF PROPERTY
INDUSTRIAL
<img file="MX339262B_D0039.tif" />
container (1) if the surfaces of the layers · clutch in a sliding or fixed way.
The amount of gas (6) that exerts a quantity of pressure (4) on the outer surface (5) of the flexible wall (2) to provide a continuously adjustable container of variable volume (1) to act on the fluid (3) contained within, it can be of any kind or type of gas (6) compatible with the outer surface (5) of the flexible wall (2) on which acts, such as an atmosphere, a mixture of gases, a mixture of gases that have selected partial pressures, a purified gas, a filtered gas, a conditioned gas, or the like. As in alternative embodiments of the invention, the amount of gas (6) can be replaced with an amount of fluidizable material capable of acting on the outer surface (5) of the flexible wall (2) to adjust the volume of the container (1), such as water, oil, or a solution.
With respect to certain embodiments of the invention, the gas (6) can exert an amount of pressure (4) on the outer surface (5) of the flexible wall (2) of between 0.0703 Kg / cm<sup>2</sup> (1 pound per square inch (psi)) up to approximately 35.2 Kg / cm<sup>2</sup> (500 pounds per square inch (psi)). As in other embodiments of the invention used for flow cytometry applications, the amount of gas (6) can exert pressure on the outer surface
- 35 IMPI
MEXICAN INSTITUTE 'Kg * =<sup>= l</sup>íS ^?<sup>5</sup>',. <, <OF THE INDUSTRIAL PROPERTY (5) of the flexible wall (2) between approximately 0.703 Kg / cm<sup>2</sup> (10 psi) and approximately 14.1 Kg / cm<sup>2</sup> (200 psi). Alternatively, the amount of gas (6) either within the gas collection element (14) or otherwise can be adjusted to generate a sufficient amount of pressure (4) on the outer surface (5) of the wall flexible (2) of the variable volume container (1) to generate a fluid stream (7) within the flow path of a conduit (8) of a microfluidic device (16) that has a fluid pressure of between 0.703 Kg / cm<sup>2</sup> (10 psi) and approximately 14.1 Kg / cm<sup>2</sup> (200 psi) or sufficient fluid pressure to generate a fluid stream (7) within the flow path of the conduit (8) that has a velocity sufficient to transport particles for a particular type or class of application, analysis, differentiation or separation.
Again, referring primarily to Figure 1A, particular embodiments of the invention may further include the fluid pressure generator (17), such as a peristaltic pump, piston pump, or the like to generate sufficient pressure for certain microfluidic or other applications. applications, in the range of approximately 7.03 Kg / cm<sup>2</sup> (100 psi) and approximately 352 Kg / cm<sup>2</sup> (5000 psi). An illustrative embodiment of the invention provides a microfluidic device (16) configured as a
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY high pressure liquid chromatography (HPLC) that has a fluid pressure generator (17) that increases the fluid pressure inside the duct (8) to between approximately 7.03 Kg / cm<sup>2</sup> (100 psi) and approximately 211 Kg / cm<sup>2</sup> (3000 psi) for applications such as normal phase or reverse phase liquid chromatography.
Now referring primarily to Figure 2A, a conventional substantially cylindrical liner fluid tank (18) (or other liner fluid tank configuration) may have an opening member (19). The opening element (19) of the coating fluid tank (18) can be configured to mate with a removably sealable lid (20) which may further include a lid securing element (21) to secure the removably sealable cap (20) . Alternative arrangements of the lid securing element (21) may include as examples, spiral threads attached to the removable sealable lid (20) and the coating fluid tank (18),
I the coiled threaded rods connected to the coating fluid tank that mate with the coiled threaded tooling that optionally applies pressure to the removably sealable cap (20), bands, retainers, or the like.
A gas inlet element (22) allows the supply of a quantity of gas (6) (various types and classes of gases as described above) to the interior of the fuel tank.
<img file="MX339262B_D0040.tif" />
- 37 IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY applications
<img file="MX339262B_D0041.tif" />
coating fluid (18).
In conventional, a quantity of fluid (3) is contained by the coating fluid tank (18) and the amount of gas (6) supplied to the interior of the coating fluid tank (18) exerts a quantity of pressure (4) on the surface of the fluid (3). A portion of the fluid (3) under pressure flows through the fluid outlet member (23) to be supplied as a fluid stream (7) in the flow path of a flow cytometer (24) (or other microfluidic device). . A pressure adjusting element (25) (such as a pressure relief valve) can allow adjustment of the amount of pressure within the coating fluid tank (18).
Referring now primarily to Figure 2B, a conventional liner fluid tank 18 (or similar fluid tank) can be adapted to operate in accordance with the invention. A variable volume container (1) having a flexible wall (2) can contain an amount of fluid (3) (coating fluid for flow cytometric applications). The variable volume container (1) containing the fluid (3) can be located inside the conventional coating fluid tank (18) by transfer through the opening element (19). A conduit (8) provides a flow path between the variable volume container
IMPI
MEXICAN INSTITUTE OF PROPERTY
INDUSTRIAL
<img file="MX339262B_D0042.tif" />
(1) and the fluid outlet element (23). A coupling element (26) may be required to connect the conduit (8) to the fluid outlet element (23) of the coating fluid tank (18). The coupling element may in certain cases comprise coupled spirally threaded tooling that operates to compress a ferrule against a seat to seal the flow path within conduit (8) against the leaking fluid. Naturally, a variety of tooling can be used since the coupling element (26) provides a continuous flow path to the fluid outlet element (23).
Now referring mainly to Figure 2C, · as in certain embodiments of the invention the variable volume container (1) can be enclosed by a second layer (12) of substantially fixed configuration as discussed above formed from a material such as a plastic, fiberglass, glass, metal, steel, polycarbonate, acrylic, polypropylene, vinyl, fluorocarbon, carbon fiber, cardboard, cardboard, or the like. The second layer can be perforated or sufficiently permeable to an amount of gas (6) to allow an amount of pressure (4) to be exerted on the outer surface (5) of the flexible wall (2) to act on the amount of the liquid (3) of the coating fluid contained inside to generate a fluid stream (7)
IMPI
MEXICAN INSTITUTE OF PROPERTY
INDUSTRIAL
<img file="MX339262B_D0043.tif" />
within the flow path of the duct (8).
Now, referring primarily to Figure 3A, certain embodiments of the invention may provide a receptacle (27) of substantially fixed configuration (rectangular as shown by Figure 3A or otherwise desired) in which one or more containers of variable volume (1) that have a flexible wall (2). The receptacle (27) can be installed on a base (28) that orients the receptacle (27) relative to a support surface (29) (eg, angled as shown in Figure 3A or substantially perpendicular to the support surface (29) as shown by Figure 3B) which can facilitate the flow of the fluid (3) inside the variable volume container (1) towards the conduit (8) that communicates with the fluid outlet element (23) . The receptacle (27) of substantially fixed configuration can be made of a material such as a plastic, a fiberglass, a glass, a metal, a steel, a polycarbonate, an acrylic, a polypropylene, a vinyl, a fluorocarbon, a fiber carbon or the like. A portion or all of the receptacle (27) can be made of a material that allows visual observation of the variable volume container (1) and the fluid (3) within the variable volume container (1). The receptacle (27) may further include a gas inlet element (22) through which a quantity can be introduced
<img file="MX339262B_D0044.tif" />
IMPI
MEXICAN INSTITUTE OF PROPERTY of gas (6) towards the gas collection element ^ TT ^ the interior surface of the receptacle (27J and T5. SUyélfieltí exterior (5) of the variable volume container (1). A pressure adjustment element ( 25) may also be included to maintain the necessary or desired amount of gas pressure (4) exerted on the outer surface (5) of the variable volume container (1).
Now, referring primarily to Figure 4A, certain embodiments of the invention may include a plurality of receptacles (27), individually spaced apart or as a single integral piece (as shown by Figures 4A and 4B) that provide a corresponding plurality of elements gas collection (14). Figure 4A illustrates that each of the plurality of gas collection elements (14) can provide independent gas inlet elements (22), fluid outlet elements (23), and pressure adjustment elements (25) to allow that each of the plurality of receptacles (27) be used independently of the other receptacles (27). In this configuration of the invention, an amount of gas (6) can be supplied to each gas collection element (14) to establish an amount of pressure (4) on the outer surface (5) of the flexible wall (2) of the variable volume container (1) located within the individual receptacle (27). Accordingly, a
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL FROMEDAD
<img file="MX339262B_D0045.tif" />
quantity of the fluid (3) contained in each of the variable volume containers (1) can be supplied to the fluid outlet element (23) from each receptacle (27). The fluid flow rate of each variable volume container (1) can be adjusted to be substantially the same or can be adjusted variably between the receptacles (27).
Now, referring mainly to Figure 4B, Alternative embodiments of the invention may be provided with a single gas inlet element (22) to supply an amount of gas (6) to all gas collection elements (14) to establish a substantially similar amount of gas pressure (4 ) on the outer surface (5) of the flexible wall (2) of each one of the plurality of containers of variable volume (1) in each corresponding one of the plurality of receptacles (27) that can be adjusted by means of a pressure adjusting element only (25). Each receptacle can further provide a fluid outlet element (23) through which a fluid stream (7) can flow into the flow path of a microfluidic device (16) (similar to that shown in Figure IA).
Now, referring mainly to Figure 5, a generic microfluidic device according to the invention is illustrated. A quantity of gas (6) can
<img file="MX339262B_D0046.tif" />
IMPI
USTITUI OR MEXICAN OF INDUSTRIAL PROPERTY supplied with a differential pressure generator (30) such as a pressurized gas tank, a gas compressor or the like, to one or more gas inlet elements (22) of the receptacle (27) through a gas transfer duct (31). A pressure regulator (32) may further be included to regulate the pressure of the amount of gas (6) in the gas transfer line (31). The amount of gas (6) is transferred from the gas inlet element (22) to a gas collection element (14) within the receptacle (27) which can have a substantially fixed configuration as shown or alternately as is described herein. At least one variable volume container (1) as described above can be located within the receptacle (27).
The amount of gas (6) within the gas collection element (14) acts on the outer surface (5) of at least one container of variable volume (1) located inside the receptacle (27) to generate a stream of fluid ( 7) in the fluid outlet element (25) that can be transferred within one or a plurality of conduits (8). The ducts (8) can have substantially the same internal diameter or vary the internal diameters. The conduit (8) may further include a fluid conditioning element (33) such as a fluid filter, a gas scrubber, or a pressure regulator.
- 43 IMPI
INSTrnJTl) MEXICANO DELAHtOHFÜAD INDUSTRIAL fluid, a fluid pressure generator, such as a pump
<img file="MX339262B_D0047.tif" />
or various permutations or combinations thereof. The conduit (8) can be connected to the flow path of a microfluidic device (24) such as a flow cytometer as shown in Figure 5 or another microfluidic device such as a fluid distribution device that transfers liquid (s) to and between locations, on a liquid confining element such as plates having a plurality of wells, the surface of the slides, specimens, channels, or other characteristics of the container.
Regarding the modality of the flow cytometer of the. invention shown in Figure 5, the variable volume container (1) having a flexible wall (2) can act on an amount of fluid (3) within the variable volume container (1) in response to an amount of pressure (4) exerted on the outer surface (5) of the flexible wall (2) by means of a quantity of gas (6) to generate the fluid stream (7) in which the particles (33) (as described above) can be transported in the supply from a source particle (34). With respect to certain flow analysis or flow classification applications, the variable volume container can be set to further allow gravity to act on the amount of fluid to aid in transfer
- 44 IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY
<img file="MX339262B_D0048.tif" />
of the quantity of fluid (3) towards the outlet element of __Lililí — jOMium ιτι-πτ * '<sup>-</sup><sup>5</sup> fluid (23). For various flow analysis and flow classification applications, the fluid stream (7) can be pressurized within a range of approximately from 15 pounds per square inch to 80 pounds per square inch as described above. With respect to certain flow classification applications, the pressure of the fluid stream (7) can be adjusted within a range from approximately 40 pounds per square inch to 50 pounds per square inch. But as an example, the fluid stream (7) can be adjusted up to about 45 pounds per square inch with a pressure variation of as little as about +/- 0.01 pounds per square inch. It may be important to maintain the fluid stream (7) at a substantially constant pressure because the consistent transport of particles (33) in the fluid stream (7) depends on keeping the pressure differential between the fluid stream substantially constant ( 7) and the source of particles (34). Unlike certain types of alternative pumps that can generate pressure fluctuations in the fluid stream (7), the invention can generate a fluid stream (7) that has constant enough pressure to in turn maintain the differential pressure between the stream d fluid (7) and the source of particles (34) to allow the transport of
IMPI
MEXICAN INDUSTRIAL PROPERTY OUTITOT
<img file="MX339262B_D0049.tif" />
particles (33) for flow analysis or for flow classification applications. Additionally, because the amount of liquid (3) in the variable volume container (1) can be protected from contaminants as described above, the constancy of the fluid stream (7) established by the invention may be greater than in flow analysis or conventional flow classification devices. The fluid stream (7) having transported particles (33) can be oscillated by a nozzle (35) to generate a plurality of drops (36) below the nozzle (35). Each of the plurality of drops (36) can carry an individual particle (33). A light source (37), such as a laser, can emit a light beam (38) or a plurality of light beams can be generated by using a beam splitting element (39) (or by using a plurality of sources (37)), which can be focused on. through an optical element (40) incident on the particle (33) transported in the fluid stream (7) below the nozzle (35), either as a single light beam or a plurality of light beams, or either at the same or different wavelengths. As for some embodiments of the invention, the characteristics of the light beam (38) can be altered by incidence on the particle (33) within the fluid stream (7) and as for other embodiments of the invention, the particle (or ligands).
IMPI MEXICAN STATUTE OF THE INDUSTRIAL EROEIEDAO
<img file="MX339262B_D0050.tif" />
fluorescent materials or the like, attached to the particle) can generate an emission (41). The light beam (s) having modified characteristics or the emission (41) can be received by a single or a plurality of detectors (42) that can generate a signal for analysis to differentiate the particles (33) transported in the drops (36) based on one or a plurality of particle characteristics. Differentiated particles can be separated based on the presence or absence of one or a plurality of particle characteristics in individual collection elements (43). The separation device (44) can include a drop charge generator (45) that induces a positive or negative charge on each drop (36) and a drop deflector (46) that acts on the charged drops to establish a path towards the appropriate collection element (43).
Now referring mainly to Figure 6, a bivariate graph generated during sperm flow sorting is shown in populations having the X chromosome and having the Y chromosome according to the invention. The bivariate graph shows that a mixture of sperm cells carrying the X chromosome and sperm cells carrying the Y chromosome can be reduced in the first population carrying the X chromosome (49) and the second population carrying the Y chromosome (50). The provision of the bivariate graph is not proposed for
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL NATURE
<img file="MX339262B_D0051.tif" />
limit yourself with respect to the numerous —- and —— waxiads. applications of the invention. Instead, the bivariate graph is intended to be illustrative of the wide range of applications in which the invention can be used. Cell flow classification can be very .... Sperm cell classification can be much more difficult than ...
Now referring primarily to Figure 7, certain embodiments of the invention can provide a plurality of variable volume containers (1) configured as a single integral piece formatted in columns and rows or otherwise as needed or desired. A plurality of receptacles (27) configured as a single integral part formatted in columns and rows can receive the plurality of containers of variable volume (1). A releasable sealable enclosure (20) can be configured to isolate each of the plurality of variable volume containers (1). A quantity of gas (6) can be supplied through a gas inlet element (22a) (22b) (two modes shown) to the gas collection element (14) within each separate receptacle (27), since either for a single receptacle of the plurality of receptacles or for a plurality of receptacles substantially simultaneously. The amount of gas (6) exerts a quantity of pressure (4) on
IMPI
MEXICAN INSTITUTE OF INDUSTRIAL PROPERTY
<img file="MX339262B_D0052.tif" />
the flexible walls (2) of the individual variable volume containers (1) to generate a fluid stream in one or a plurality of conduits (8) that communicate with each receptacle (27).
Now referring primarily to Figure 8, the conduit (8) that fluidly communicates with each receptacle (27) may comprise a microfluidic conduit (inner diameter of one millimeter or less) such as a plastic tube, or as shown in Figure 8 it may also comprise a discharge element (44) on the surface of a single or a plurality of fluid supply bodies (45) that provides a flow path for the fluid stream (7). Fluid supply bodies 45 can be releasably and interchangeably sealed to provide a number of different flow paths. In the embodiment shown, the flow path established by the releasably sealable fluid supply bodies can supply fluid (3) from a plurality of variable volume containers (1) to a plate (46) having a plurality of wells (47).
V. EXAMPLES
EXAMPLE 1. Referring now to Figure 8, which shows a bivariate graph generated from analysis of differentiated fluorochrome stained sperm cells
IMPI
MEXICAN INSTITUTE OF PROPERTY
INDUSTRIAL
<img file="MX339262B_D0053.tif" />
based on the presence of a r.rnmnsnm ^ xo nn ..v.
using a MoFlo® flow cytometer from DakoCytomation, Inc., according to the invention. A conventional liner fluid tank is modernized with a variable volume container according to the invention containing approximately 5 liters of sterile liner fluid. The coating fluid was kept at about 20 ° C during use. An amount of gas was supplied to the coating fluid tank to exert an amount of gas pressure on the outer surface of the variable volume container resulting in the generation of a fluid stream within the flow path of a flow cytometer MoFlo® from DakoCytomation, Inc. The flow cytometer was then otherwise operated according to standard operating procedures provided by DakoCytomation, Inc., for a period of approximately 8 hours to analyze and classify a mixture of sperm cells to generate a viable population of sperm carrying the X chromosome and a viable population of sperm that carry the Y chromosome. Enriched populations carrying the X chromosome and carrying the Y chromosome were established in separate collection containers.
EXAMPLE 2. Similarly, a flow cytometer that classifies human sperm according to the invention,
IMPI
MEXICAN INSTITUTE Say THE INDUSTRIAL PROPERTY
<img file="MX339262B_D0054.tif" />
carry the X chromosome and insemination purpose
Sperm cells
- 50 can provide populations that carry the Y chromosome for the artificial selected by sex, human sufficient for the artificial insemination of a human female can be classified by flow in approximately 2 hours from the ejaculation of male human. Enriched populations of human sperm carrying the X chromosome or carrying the Y chromosome may be more than 80% pure. Clinical procedures may require that after each sample is classified, the fluidic classification channels are washed with an acid wash, a base wash, a disinfecting wash, and then a water wash. The present invention can be used to supply four different sterile fluids to the flow cytometer and allow automated computer cleaning steps to be carried out between patients. During the automated washing procedure, the doctor can carry out the artificial insemination procedure.
EXAMPLE 3. According to the invention, a plurality of different microfluidic devices can be operated 24 hours a day. Variable volume containers can be placed in a common pressurized receptacle at approximately 1.6 atmospheres. Each microfluidic device can be served with one or more conduits from
INDUSTRIAL
IMPI
INSTITUTO MEXICANO de la reonBwn
<img file="MX339262B_D0055.tif" />
of variable volume containers that corniuiii- litl <sup>iww</sup> ·<sup>λΊ</sup>'~ * ~ Conventional tooling of the microfluidic device.
As can be easily understood from the foregoing, the basic concepts of the present invention can be incorporated in a variety of ways. The invention involves numerous and diverse embodiments of a continuously variable volume container for supplying fluid and methods for making and using such a continuously variable volume container.
As such, the particular embodiments or elements of the invention described by the description or shown in the figures accompanying this application are not intended to be limiting, but more so. either exemplary of the numerous and varied modalities encompassed generically by the invention or equivalents covered with respect to any particular element thereof. Furthermore, the specific description of a single embodiment or element of the invention may not explicitly describe all possible embodiments or elements; many alternatives are implicitly described by the description and the figures.
It should be understood that each element of an apparatus or each step of a method can be described by an apparatus term or method term. Such terms can be substituted where you want to make the coverage explicit.
<img file="MX339262B_D0056.tif" />
implicitly broad to which you have right
<img file="MX339262B_D0057.tif" />
But as an example, it should be understood that all stages of a method can be described as an action, a means of taking that action, or as an element causing that action. Similarly, each element of an apparatus can be described as the physical element or the action that facilitates that physical element. But as an example, the description of an adjustable volume should be understood as encompassing the description of the act of adjusting the volume - whether explicitly or not - and conversely, where the act of adjusting the volume is effectively described , such a description is to be understood to encompass the description of an adjustable volume and still a means of adjusting the volume. Such alternative terms for each element or stage should be understood to be explicitly included in the description.
Furthermore, as for each term used it should be understood that unless their use in this application is inconsistent with such an interpretation, common dictionary definitions should be understood to be included in the description for each term as contained in the Random House Webster<sup>z</sup>s Unabridged Dictionary, second edition, each definition incorporated herein by reference.
Thus, it should be understood that the applicant (s) ί ΪΜΡΙ
Instituto Mexicano OE LA FRONEDAD INDUSTRIAL
<img file="MX339262B_D0058.tif" />
claims (n) at least: i) each of the fluid supply devices discussed and described herein, ii) the related methods discussed and described, iii) similar, equivalent and even implicit variations of each of these devices and methods, iv) those alternative modalities that carry out each of the functions shown, treated or described, v) those alternative designs and methods that perform each of the functions shown as implicit to carry out what is discussed and described, vi) each characteristic, component and stage shown as separate and independent inventions, vii) the applications improved by the various systems or components described, viii) the resulting products produced by such systems or components, ix) methods and apparatus substantially as described herein and with reference to any of the accompanying examples, x) the various combinations and permutations of each of the described pre-elements.
The claims set forth in this specification are incorporated herein by reference as part of this description of the invention and the applicant expressly reserves the right to use all or a portion of such embedded content of such claims as further description to support any or all claims or any
IMPI c / ι INSTITUTO MEXICANO ““ DE LA PROPIEDAD
INDUSTRIAL element or component thereof and the applicant furthermore expressly reserves the right to withdraw any portion or all of the incorporated content of such claims or any element or component thereof from the description in the claims or vice-versa as necessary to define the subject matter for which protection is sought through this request or through any continuation request, division or continuation in a subsequent part of the same or to obtain any benefit, reduction of rights, or to comply with the laws, rules or regulations of patent of any country or treaty and such content incorporated by reference will survive throughout the processing of this request including any request for continuation, division or continuation in a subsequent part thereof or any reissue or extension thereof.
The claims set forth below are intended to describe the scopes and limits of a limited number of the preferred embodiments of the invention and are not to be construed as the broader embodiment of the invention or a complete listing of the embodiments of the invention that can be claimed. The applicant does not waive any right to develop additional claims based on the description set forth above as part of any request for continuation, division or continuation
<img file="MX339262B_D0059.tif" />
IMPI
INSTITUTO MEXICANO DELA PROPIEDAD INDUSTRIAL in part or the like.
-561.
to
Contents92
67 sheets
Sheet 1 Sheet 2 Sheet 3 Sheet 4 Sheet 5 Sheet 6 Sheet 7 Sheet 8 Sheet 9 Sheet 10 Sheet 11 Sheet 12 Sheet 13 Sheet 14 Sheet 15 Sheet 16 Sheet 17 Sheet 18 Sheet 19 Sheet 20 Sheet 21 Sheet 22 Sheet 23 Sheet 24 Sheet 25 Sheet 26 Sheet 27 Sheet 28 Sheet 29 Sheet 30 Sheet 31 Sheet 32 Sheet 33 Sheet 34 Sheet 35 Sheet 36 Sheet 37 Sheet 38 Sheet 39 Sheet 40 Sheet 41 Sheet 42 Sheet 43 Sheet 44 Sheet 45 Sheet 46 Sheet 47 Sheet 48 Sheet 49 Sheet 50 Sheet 51 Sheet 52 Sheet 53 Sheet 54 Sheet 55 Sheet 56 Sheet 57 Sheet 58 Sheet 59 Sheet 60 Sheet 61 Sheet 62 Sheet 63 Sheet 64 Sheet 65 Sheet 66 Sheet 67
27 members in 11 offices
Priority claims9
| Document | Office | Kind | Date |
|---|---|---|---|
| 11004382 | United States of America | – | |
| 438204 | United States of America | A | |
| 438204 | United States of America | A | |
| 2005043926 | United States of America | W | |
| 2005043926 | United States of America | W | |
| 11004382 | – | – | – |
| US0543926 | – | – | – |
| US20040004382 | – | – | – |
| WO2005US43926 | – | – | – |
Members27
| Document | Office | Kind | |
|---|---|---|---|
| AU2005311618A1 | Australia | A1 | |
| CA2590191A1 | Canada | A1 | |
| US2006118167A1 | United States of America | A1 | |
| WO2006060770A2 | World Intellectual Property Organization (WIPO) | A2 | |
| WO2006060770A3 | World Intellectual Property Organization (WIPO) | A3 | |
| MX2007006590A | Mexico | A | |
| EP1834167A2 | European Patent Office (EPO) | A2 | |
| CN101111756A | China | A | |
| BRPI0518598A2 | Brazil | A2 | |
| AU2005311618B2 | Australia | B2 | |
| NZ555801A | New Zealand | A | |
| US2011000934A1 | United States of America | A1 | |
| CN101111756B | China | B | |
| US8080422B2 | United States of America | B2 | |
| US2012107805A1 | United States of America | A1 | |
| EP1834167A4 | European Patent Office (EPO) | A4 | |
| MX339262BThis record | Mexico | B | |
| EP3236233A1 | European Patent Office (EPO) | A1 | |
| EP1834167B1 | European Patent Office (EPO) | B1 | |
| BRPI0518598B1 | Brazil | B1 | |
| DK1834167T3 | Denmark | T3 | |
| HUE038869T2 | Hungary | T2 | |
| US10190964B2 | United States of America | B2 | |
| US2019204209A1 | United States of America | A1 | |
| CA2590191C | Canada | C | |
| US11175213B2 | United States of America | B2 | |
| EP3236233B1 | European Patent Office (EPO) | B1 |
Numbers
- Publication
- 339262
- Publication, DOCDB
- 339262
- Publication, EPODOC
- MX339262
- Application
- 2011007610
- Application, DOCDB
- 2011007610
- Application, EPODOC
- MX20110007610
Titles
- Spanish
- CONTENEDOR DE VOLUMEN CONSTANTEMENTE VARIABLE REGULADO POR PRESION PARA EL SUMINISTRO DE FLUIDO.
Classification
- CPC, 17
- G01N15/1484
- B01L3/0296
- B01L3/5027
- B01L2200/027
- B01L2200/0684
- B01L2200/141
- B01L2400/0481
- G01N35/1095
- Y10T436/117497
- Y10T436/11
- Y10T436/2575
- Y10T436/118339
- Y10T137/0318
- Y10T137/1624
- G01N15/1409
- G01N15/1492
- G01N15/149
- IPC, 3
- B01L3 00
- F16K17 40
- G01N35 08
