Ribbon flow cytometry and cell sorting

• US 20030175980A1
• Filed: 02/20/2003
• Published: 09/18/2003
• Est. Priority Date: 03/14/2002
• Status: Abandoned Application

• Current Assignee: Perkinelmer Health Sciences Incorporated (Perkinelmer Incorporated)
• Original Assignee: Perkinelmer Health Sciences Incorporated (Perkinelmer Incorporated)
• Inventors: Hayenga, Jon W., Weigl, Bernhard H., Bardell, Ronald L.
• Application Number:
  US10/370,902
• Publication Number:
  US 20030175980A1
• Time in Patent Office: Days
• US Class Current: 436/63
• 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 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0622 distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• 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: B07C 5/342 according to optical properties, e.g. colour according to radiation transmittivity B07C5/3416
• CPC Class Codes: B07C 5/344 according to electric or electromagnetic properties sorting according to size measured electrically or electronically B07C5/08; material testing by magnetic means G01N24/00, G01N27/00, by electrical means G01N27/00; electrical measuring devices in general G01R; coin testing G07D5/00
• CPC Class Codes: G01N 15/1404 Handling flow, e.g. hydrodynamic focusing
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/1484 microstructural devices
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1413 Hydrodynamic focussing

Abstract

A microfluidic device for sorting cells is described. The device includes an input channel, a primary channel, at least two branch channels which connect with the primary channel at a junction, and a sheath injector positioned upstream from the junction. Sample solution, which may contain a population of cells, can be entered into the input channel and hydrodynamically focused into a sample ribbon. The device employs a system for directing fluid flow, and particularly the flow of the sample ribbon, into a branch channel based on a detected cell feature. The system can employ a variety of sorting techniques to change or direct the flow of cells into a particular branch channel.

Claims

1. A microfluidic device for sorting cells, comprising:

a. an input channel for receiving sample solution;

b. a primary channel, with a first end of the primary channel connected with a sheath solution reservoir and a second end connected with at least two branch channels which connect at a junction;

c. a sheath injector positioned upstream from the junction and connected with the input channel such that sample solution entering the primary channel flows through the sheath injector and is hydrodynamically focused into a ribbon compressed between at least two layers of sheath solution;

d. an interrogation region positioned between the junction and the sheath injector, where the interrogation region is coupled with a recognition apparatus for evaluating the cells in the ribbon according to at least one cell feature as the ribbon passes through the interrogation region; and

e. a cell diversion system responsive to the recognition apparatus and capable of directing fluid flow into a branch channel based on a detected feature.

2. The microfluidic device of claim 1, wherein the cell diversion system consists of at least one valve positioned along each branch channel, with no more than the valves of one branch channel being open at any one time, and the status of the valves being determined through communication with the recognition apparatus.

3. The microfluidic device of claim 1, wherein the cell diversion system consists of a means for manipulating pressure within the branch channels, such that flow within the branch channels is differentially restricted and the sample ribbon can be directed into a branch channel.

4. The microfluidic device of claim 1, wherein the cell diversion system consists of a channel for introducing fluid, said channel being positioned upstream from the junction, with introduction of fluid from said channel being determined through communication with the recognition apparatus.

5. The microfluidic device of claim 1, wherein the cell diversion system consists of a channel for removing fluid from the primary channel, said channel being positioned upstream from the junction, with introduction of fluid from said channel being determined through communication with the recognition apparatus.

6. The microfluidic device of claim 1, wherein the cell diversion system consists of a magnetic source positioned proximate to the junction, such that cells conjugated with a label having a magnetically responsive portion and flowing through the junction are directed into one or more branch channels in response to the magnetic source.

7. The microfluidic device of claim 1, wherein the cell diversion system consists of an applied electrical field positioned proximate to the junction, such that cells flowing through the electrical field are directed into one or more branch channels in response to the interaction between the charge of the cell and the applied electrical field.

8. The microfluidic device of claim 1, further comprising a means for geometrically focusing the sample ribbon.

9. The microfluidic device of claim 1, wherein each evaluated cell feature is made detectable by use of a label.

10. The microfluidic device of claim 1, where labeled cells emit a fluorescent signal, which is detected by the recognition apparatus consisting of an optical detection means.

11. A sorting unit contained within a cartridge, comprising:

a. an input channel for receiving sample solution;

b. a primary channel, with a first end of the primary channel connected with a sheath solution reservoir and a second end connected with at least two branch channels which connect at a junction;

c. a sheath injector positioned upstream from the junction and connected with the input channel such that sample solution entering the primary channel flows through the sheath injector and is hydrodynamically focused into a ribbon compressed between at least two layers of sheath solution;

d. an interrogation region positioned between the junction and the sheath injector, where the interrogation region is coupled with a recognition apparatus for detecting labeled cells as the ribbon passes through the interrogation region; and

e. a cell diversion system responsive to the recognition apparatus and capable of directing fluid flow into a branch channel based on a detected feature.

12. The sorting unit of claim 11, wherein the cell diversion system consists of at least one valve positioned along each branch channel, with no more than the valves of one branch channel being open at any one time, and the status of the valves being determined through communication with the recognition apparatus.

13. The sorting unit of claim 11, wherein the cell diversion system consists of a means for manipulating pressure within the branch channels, such that flow within the branch channels is differentially restricted and the sample ribbon can be directed into a branch channel.

14. The sorting unit of claim 11, wherein the cell diversion system consists of a channel for introducing fluid, said channel being positioned upstream from the junction, with introduction of fluid from said channel being determined through communication with the recognition apparatus.

15. The sorting unit of claim 11, wherein the cell diversion system consists of a channel for removing fluid from the primary channel, said channel being positioned upstream from the junction, with introduction of fluid from said channel being determined through communication with the recognition apparatus.

16. The sorting unit of claim 11, wherein the cell diversion system consists of a magnetic source positioned proximate to the junction, such that cells conjugated with a label having a magnetically responsive portion and flowing through the junction are directed into one or more branch channels in response to the magnetic source.

17. The sorting unit of claim 11, wherein the cell diversion system consists of an applied electrical field positioned proximate to the junction, such that cells flowing through the electrical field are directed into one or more branch channels in response to the interaction between the charge of the cell and the applied electrical field.

18. The sorting unit of claim 11, further comprising a means for geometrically focusing the sample ribbon upstream from the interrogation region.

19. A method of sorting cells in a microfluidic device, comprising the steps of:

a. hydrodynamically focusing a population of cells into a ribbon compressed between at least two layers of sheath solution;

b. flowing the ribbon through a channel comprising an interrogation region;

c. determining the presence or amount of label on each cell as it passes through the interrogation region; and

d. diverting cells into a first branch channel or a second branch channel based on the presence or amount of label on each cell.

20. The method of claim 19, further comprising the step of geometrically focusing the ribbon upstream from the sample ribbon.

21. The method of claim 19, wherein the cell diverting step includes positioning at least one valve along each branch channel, such that no more than the valves of one branch channel are capable of being open at any one time, the status of the valves being determined through communication with the recognition apparatus.

22. The method of claim 19, wherein the cell diverting step includes means for manipulating pressure within the branch channels, such that flow within the branch channels is differentially restricted and the sample ribbon can be directed into a branch channel.

23. The method of claim 19, wherein the cell diverting step includes positioning a channel for introducing fluid upstream from the junction, with introduction of fluid from said channel being determined through communication with the recognition apparatus.

24. The method of claim 19, wherein the cell diverting step includes positioning a channel for removing fluid from the primary channel upstream from the junction, with introduction of fluid from said channel being determined through communication with the recognition apparatus.

25. The method of claim 19, wherein the cell diverting step includes positioning a magnetic source proximate the junction, such that cells conjugated with a label having a magnetically responsive portion and flowing through the junction are directed into one or more branch channels in response to the magnetic source.

26. The method of claim 19, wherein the cell diverting step includes positioning an applied electrical field proximate to the junction, such that cells flowing through the electrical field are directed into one or more branch channels in response to the interaction between the charge of the cell and the applied electrical field.