Method and apparatus for sorting particles

• US 20070065808A1
• Filed: 11/22/2006
• Published: 03/22/2007
• Est. Priority Date: 04/17/2002
• Status: Abandoned Application

• Current Assignee: Cytonome/St, Llc (Cytonome Inc.)
• Original Assignee: Cytonome Inc.
• Inventors: Deshpande, Manish, Bohm, Sebastian, Gilbert, John
• Application Number:
  US11/603,444
• Publication Number:
  US 20070065808A1
• Time in Patent Office: Days
• US Class Current: 435/4
• CPC Class Codes: B01L 2200/0636 Focussing flows, e.g. to laminate flows
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/502776 specially adapted for focusing or laminating flows
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties

Abstract

A method and apparatus for sorting particles moving through a closed channel system of capillary size comprises a bubble valve for selectively generating a pressure pulse to separate a particle having a predetermined characteristic from a stream of particles. The particle sorting system may further include a buffer for absorbing the pressure pulse. The particle sorting system may include a plurality of closely coupled sorting modules which are combined to further increase the sorting rate. The particle sorting system may comprise a multi-stage sorting device for serially sorting streams of particles, in order to decrease the error rate.

Claims

1. A method of isolating a subpopulation of cells from a cell population using a microfluidic cell sorting device comprising;

identifying cells from a population in a mixture that have a desired phenotype;

isolating said cells from cells that do not have the desired phenotype using a microfluidic cell sorting device;

passing the isolated cells having the desired phenotype to a mixing and incubation region of the microfluidic cell sorting device; and

introducing a test compound to the mixing and incubation region.

2. The method of claim 1, further comprising the step of:

detecting the effect of the test compound on the isolated cells having the desired phenotype in a detection region of the microfluidic cell sorting device.

3. The method of claim 1 wherein the step of isolating said cells comprises the steps of:

conveying the mixture including cells having the desired phenotype and cells that do not have the desired phenotype through a sorting channel of the microfluidic cell sorting devices,

using an actuator in the microfluidic cell sorting device to apply a pressure pulse to a cell having the desired phenotype to deflect the cell having the desired phenotype into a first outlet of the sorting channel while cells not having the desired phenotype flow into a second outlet of the sorting channel, absorbing the pressure pulse in the sorting channel using a buffer,

4. A method of isolating a subpopulation of cells in a mixture comprising,

identifying a subpopulation of cells in the mixture displaying a cell cycle stage specific marker;

isolating said subpopulation of cells using a microfluidic cell sorting device;

passing said subpopulation of cells that are in the same phase of the cell cycle to a mixing and incubation region in the microfluidic cell sorting device; and

introducing a test compound to the mixing and incubation region.

5. The method of claim 4, wherein the step of isolating said subpopulation of cells comprises the steps of:

conveying the mixture including cells having the desired phenotype and cells that do not have the desired phenotype through a sorting channel of the microfluidic cell sorting device,

using an actuator in the microfluidic cell sorting device to apply a pressure pulse to a cell having the desired phenotype to deflect the cell having the desired phenotype into a first outlet of the sorting channel while cells not having the desired phenotype flow into a second outlet of the sorting channel, and

absorbing the pressure pulse in the sorting channel using a buffer;

thereby isolating a subpopulation of cells that are in the same phase of the cell cycle.

6. A method of isolating a subpopulation of cells from a cell population using a system comprising;

identifying cells from a population that have a desired phenotype;

measuring a velocity of a cell having the desired phenotype; and

isolating said cells from cells that do not have the desired phenotype using the microfluidic cell sorting device based on the steps of identifying cells having the desired phenotype and the measuring the velocity.

7. A method of isolating a subpopulation of cells from a cell population in a mixture using a microfluidic cell sorting device comprising the steps of:

conveying a mixture including cells having a desired phenotype and cells that do not have the desired phenotype through a sorting channel of the microfluidic cell sorting device;

identifying cells from a population of cells in the mixture that have a desired phenotype; and

using an actuator in the microfluidic cell sorting device to deflecting a meniscus formed by fluid at an intersection between a side channel in communication with the sorting channel and a sealed chamber positioned adjacent to the side channel to apply a pressure pulse to a cell having the desired phenotype to deflect the cell having the desired phenotype into a first outlet of the sorting channel while cells not having the desired phenotype flow into a second outlet of the sorting channel, thereby isolating said cells from cells that do not have the desired phenotype using a microfluidic cell sorting device.

8. The method of claim 7, wherein the cells having the desired phenotype are cells to be used in transplantation.

9. The method of claim 7, wherein the cells having the desired phenotype are cells to be genetically modified.

10. The method of claim 7, further comprising the step of measuring a velocity of a cell having the desired phenotype.

11. The method of claim 7, further comprising the step of genetically modifying the cells having the desired phenotype.

12. The method of claim 11, further comprising the step of transplanting the isolated cells containing the desired phenotype.