Rapid Single Cell Based Parallel Biological Cell Sorter

• US 20120142018A1
• Filed: 12/01/2011
• Published: 06/07/2012
• Est. Priority Date: 12/05/2010
• Status: Active Grant

• Current Assignee: Cytek Biosciences Inc
• Original Assignee: Wenbin Jiang
• Inventors: Jiang, Wenbin
• Granted Patent: US 9,394,511 B2
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: G01N 15/01 specially adapted for biological cells, e.g. blood cells investigating sedimentation of particle suspensions in blood G01N15/05
• CPC Class Codes: G01N 15/1434 Optical arrangements
• 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 2015/1006 for cytology
• CPC Class Codes: G01N 2015/144 Imaging characterised by its optical setup

Abstract

A disposable rapid cell sorter comprises of microfluidic chip with electrodes and sorts biological cells of interest through magnetic field and electric field based on biological cell functional antibody bonded magnetic beads and luminescent labeling.

Claims

1. A rapid parallel single cell sorter comprising:

a microfluidic chip with an inlet for receiving fluid containing biological cells to be sorted, a transparent flat hollow cavity for holding the said fluid for cell sorting, and an outlet for releasing the said fluid containing the sorted cells;

a magnetic field generator to sort magnetically labeled biological cells, said magnetically labeled biological cells are cells bonded to antibodies containing magnetic micro-beads; and

a fluidic switch interfacing with the outlet to switch fluid containing the sorted cells from the outlet toward one or more sorted cell collection ports.

2. The magnetic generator in claim 1 is an array of micro-magnetics.

3. The rapid parallel cell sorter in claim 1 further comprises:

a layer of film on one of the flat hollow cavity surfaces of the said microfluidic chip;

a light source for the excitation of photoluminescence labeled biological cells; and

an optical detection system for capturing the photoluminescence from specific labeled biological cells.

4. The layer of film in claim 3 is an array of conductive electrodes for generating two-dimensionally addressable electric field extending into the microfluidic cavity according to the array pattern configuration.

5. The layer of film in claim 3 is a layer of optic cross-linkable molecule or polymer for bonding or debonding labeled biological cells to the microfluidic surface through light illumination.

6. The optical detection system in claim 3 is wavelength sensitive.

7. A rapid parallel single cell sorter comprising:

a microfluidic chip with an inlet for receiving fluid containing biological cells to be sorted, a transparent flat hollow cavity for holding the said fluid for cell sorting, and an outlet for releasing the said fluid containing the sorted cells;

a fluidic switch interfacing with the outlet to switch fluid containing the sorted cells from the outlet toward one or more sorted cell collection ports;

a layer of film on one of the flat hollow cavity surfaces of the said microfluidic chip;

a light source for the excitation of photoluminescence labeled biological cells; and

an optical detection system for capturing the photoluminescence from specific labeled biological cells.

8. The layer of film in claim 7 is an array of conductive electrodes for generating two-dimensionally addressable electric field extending into the microfluidic cavity according to the array pattern configuration.

9. The layer of film in claim 7 is a layer of optic cross-linkable molecule or polymer for bonding or debonding labeled biological cells to the microfluidic surface through light illumination.

10. The optical detection system in claim 7 is wavelength sensitive.

11. A method for rapidly sorting biological cells comprising:

magnetically label biological cells of interest using antibodies bonded with magnetic micro-bead;

receive fluid containing the biological cells to be sorted through an inlet of a microfluidic chip;

hold the said fluid for cell sorting in a transparent hollow cavity of the said microfluidic chip;

turn-on magnetic field to sort biological cells in the microfluidic chip cavity by aligning the magnetically labeled biological cells in the fluid against one of the flat surfaces of the said microfluidic chip inside the cavity;

release fluid with sorted cells not magnetically labeled through an outlet of the said microfluidic chip; and

collect the sorted cells not magnetically labeled into a first cell collection port through a fluid switch.

12. The biologic cells magnetically labeled in claim 11 are one or all of CD45 cells, CD4 cells and CD 8 cells.

13. The method in claim 11 further comprising:

turn off magnetic field and flush the sorted magnetically labeled cells leaning against the flat surface of the said microfluidic chip into a second collection port through the outlet of the microfluidic chip and the fluid switch.

14. A method for rapidly sorting biological cells comprising:

magnetically label biological cells of interest using antibodies bonded with magnetic micro-beads;

label a subset of the magnetically labeled biological cells with antibodies bonded with a specific photoluminescence label;

receive fluid containing the biological cells to be sorted through an inlet of a microfluidic chip;

hold the said fluid for cell sorting in a transparent hollow cavity of the said microfluidic chip;

turn-on magnetic field to sort biological cells in the microfluidic chip cavity by aligning the magnetically labeled biological cells in the fluid against one of the flat surfaces of the said microfluidic chip inside the cavity;

excite specific photoluminescence from the subset magnetically aligned biologic cells using a light source;

detect the photoluminescence using a detection system; and

turn on 2-dimensionally addressable electric field closest to the subset cells detected by the luminescence detection system to catch the subset cells through electric force applied to the magnetic micro-beads.

15. The method in claim 14 further comprising:

release fluid with sorted cells not magnetically labeled through an outlet of the said microfluidic chip; and

collect the sorted cells not magnetically labeled into a first cell collection port through a fluid switch.

16. The method in claim 15 further comprising:

turn off magnetic field and release sorted magnetically labeled biologic cells not electrically caught onto the microfluidic chip surface through the outlet and the fluid switch toward a second cell collection port;

17. The method in claim 16 further comprising:

turn off electric field and release sorted luminescence labeled subset biologic cells through the outlet and fluid switch toward a third collection port.

18. The microfluidic chip in claim 14 is made of one of glass, molded plastics, gel, GaN and sapphire with an array of conductive electrodes on one of the flat hollow cavity surfaces of the said microfluidic chip for generating two-dimensionally addressable electric field extending into the microfluidic cavity according to the array electrode configuration.

19. The biologic cells magnetically labeled in claim 14 are CD45 cells.

20. The subset biologic cells both magnetically and photo luminescence labeled in claim 14 are CD4 and/or CD8 cells.

21. A method for rapidly sorting biological cells comprising:

magnetically label biological cells of interest using antibodies bonded with magnetic micro-beads;

label more than one subset of the magnetically labeled biological cells with more than one type of antibodies, each antibody bonded with a specific photoluminescence label;

receive fluid containing the biological cells to be sorted through an inlet of a microfluidic chip;

hold the said fluid for cell sorting in a transparent hollow cavity of the said microfluidic chip;

turn-on magnetic field to sort biological cells in the microfluidic chip cavity by aligning the magnetically labeled biological cells in the fluid against one of the flat surfaces of the said microfluidic chip inside the cavity;

excite specific photoluminescences of different wavelengths from the more than one subset of the photo luminescence labeled biologic cells using a light source across the aligned magnetically labeled biologic cells;

detect the photoluminescence of different wavelengths using a wavelength sensitive detection system; and

turn on 2-dimensionally addressable electric field closest to the subset cells detected by the luminescence detection system to catch the subset cells through electric force applied to the magnetic micro-beads;

record the subset cell map corresponding to the photo luminescence wavelength into a memory.

22. The wavelength sensitive detection system in claim 21 is one of a CCD camera with color filters and a monochrometer.

23. The method in claim 21 further comprising:

release fluid with sorted cells not magnetically labeled through an outlet of the said microfluidic chip; and

collect the sorted cells not magnetically labeled into a first cell collection port through a fluid switch.

24. The method in 23 further comprising:

turn off magnetic field and release sorted magnetically labeled biologic cells not electrically caught onto the microfluidic chip surface through the outlet and the fluid switch toward a second cell collection port;

25. The method in claim 24 further comprising:

turn off electric field corresponding to a specific photo luminescence wavelength according to the memory map and release the correspondent sorted luminescence labeled subset biologic cells through the outlet and the fluid switch toward one of a third collection port, a fourth collection port, and thereafter.

26. A method for rapidly sorting biological cells comprising:

receive fluid containing biological cells to be sorted through an inlet of a microfluidic chip;

hold the said fluid for cell sorting in a transparent hollow cavity of the said microfluidic chip;

immobilize the said biological cells towards one of the flat surfaces of the said microfluidic chip inside the cavity using a first means;

characterize the immobilized biological cells using a second means; and

reinforce the immobilized cells using a third means according to the characterization results.

27. The first means in claim 26 comprising:

pre-label the said biological cells to be sorted with magnetic beads and immobilize the magnetic labeled cells using magnetic field.

28. The second means in claim 26 comprising:

pre-label the said biological cells to be sorted with photoluminescence labels;

excite specific photoluminescences from the photo luminescence labeled biologic cells using a light source across the immobilized biologic cells; and

detect the photoluminescence with a detection system.

29. The third means in claim 26 comprising:

apply 2-dimmensional micro-electrodes to the said flat surface of the microfluidic chip for the generation of electric field penetrating into the microfluidic chip;

turn-on the electric field to apply electric force to reinforce the immobilization of the biologic cells at the location of the electrode according to the said characterization results.