Devices and method for enrichment and alteration of cells and other particles

Patent metadata

Field	Value
Publication number	US-9956562-B2
Application number	US-201514930313-A
Country	US
Kind code	B2
Filing date	Nov 2, 2015
Priority date	Apr 5, 2005
Publication date	May 1, 2018
Grant date	May 1, 2018

Abstract

The invention features devices and methods for the deterministic separation of particles. Exemplary methods include the enrichment of a sample in a desired particle or the alteration of a desired particle in the device. The devices and methods are advantageously employed to enrich for rare cells, e.g., fetal cells, present in a sample, e.g., maternal blood and rare cell components, e.g., fetal cell nuclei. The invention further provides a method for preferentially lysing cells of interest in a sample, e.g., to extract clinical information from a cellular component, e.g., a nucleus, of the cells of interest. In general, the method employs differential lysis between the cells of interest and other cells (e.g., other nucleated cells) in the sample.

First claim

What is claimed is:

1. A method of producing a sample enriched in particles of a specific type from a liquid sample, the method comprising
obtaining a liquid sample comprising particles of the specific type;
increasing a hydrodynamic size of particles of the specific type in the liquid sample by adding to the liquid sample a reagent that swells the particles of the specific type; and
flowing the liquid sample through a channel under laminar flow conditions, wherein the channel comprises an inlet, an array of obstacles that form a network of gaps, and first and second outlets, wherein any particles in the liquid sample passing into the inlet and through the network of gaps are divided unequally into a major flux and a minor flux so that particles having a hydrodynamic size above a critical size are displaced laterally in the network of gaps to the first outlet and particles having a hydrodynamic size below the critical size are not displaced laterally in the network of gaps and flow to the second outlet, wherein the specific type of particles having an altered hydrodynamic size are directed selectively to either the first outlet or the second outlet, thereby producing a sample enriched in particles of the specific type.

2. The method of claim 1 , further comprising separating particles of the specific type from the enriched liquid sample.

3. The method of claim 1 , wherein the reagent comprises a hypotonic solution.

4. A method of producing a sample enriched in particles of a specific type from a liquid sample, the method comprising
obtaining a liquid sample comprising particles of the specific type;
decreasing a hydrodynamic size of the particles of the specific type in the liquid sample; and
flowing the liquid sample through a channel under laminar flow conditions, wherein the channel comprises an inlet, an array of obstacles that form a network of gaps, and first and second outlets, wherein any particles in the liquid sample passing into the inlet and through the network of gaps are divided unequally into a major flux and a minor flux so that particles having a hydrodynamic size above a critical size are displaced laterally in the network of gaps to the first outlet and particles having a hydrodynamic size below the critical size are not displaced laterally in the network of gaps and flow to the second outlet, wherein the specific type of particles having an altered hydrodynamic size are directed selectively to either the first outlet or the second outlet, thereby producing a sample enriched in particles of the specific type.

5. The method of claim 4 , wherein decreasing the hydrodynamic size comprises adding to the liquid sample a hypertonic solution that shrinks the particles of the specific type.

6. The method of claim 4 , further comprising separating particles of the specific type from the enriched liquid sample.

7. A method of producing a sample enriched in particles of a specific type from a liquid sample, the method comprising
obtaining a liquid sample comprising particles of the specific type;
altering a hydrodynamic size of particles of the specific type in the liquid sample; and
flowing the liquid sample through a channel under laminar flow conditions, wherein the channel comprises an inlet, an array of obstacles that form a network of gaps, and first and second outlets, wherein the channel is bounded by two side walls and further comprises a bypass channel defined by a region within the channel devoid of obstacles between a side wall and the array of obstacles, wherein any particles in the liquid sample passing into the inlet and through the network of gaps are divided unequally into a major flux and a minor flux so that particles having a hydrodynamic size above a critical size are displaced laterally in the network of gaps toward the bypass channel and to the first outlet and particles having a hydrodynamic size below the critical size are not displaced laterally in the network of gaps and flow to the second outlet, and wherein the specific type of particles having an altered hydrodynamic size are directed selectively to either the first outlet or the second outlet, thereby producing a sample enriched in particles of the specific type.

8. The method of claim 7 , wherein altering the hydrodynamic size comprises increasing the hydrodynamic size of the particles of the specific type.

9. The method of claim 8 , wherein increasing the hydrodynamic size comprises adding to the liquid sample microparticles that bind selectively to the particles of the specific type.

10. The method of claim 9 , wherein the microparticles comprise an antibody or one of an avidin-biotin binding ligand pair to bind selectively to the particles of the specific type.

11. The method of claim 10 , wherein the microparticles are magnetic or polymer beads.

12. The method of claim 8 , wherein increasing the hydrodynamic size comprises adding to the liquid sample a reagent that swells the particles of the specific type.

13. The method of claim 12 , wherein the reagent comprises a hypotonic solution.

14. The method of claim 7 , wherein the particles of the specific type are rare cells.

15. The method of claim 14 , wherein the rare cell is a blood cell, a fetal cell, a nucleated red blood cell, a stem cell, a cancer cell, an immune system cell, an epithelial cell, a connective tissue cell, a bacterial cell, a fungal cell, or a parasite.

16. The method of claim 14 , wherein the rare cells are blood cells, wherein the channel comprises a second array of obstacles that form a network of gaps and a third outlet, and wherein the method produces three separate sample fractions, wherein a first fraction is enriched in platelets, a second fraction is enriched in red blood cells, and a third fraction is enriched in white blood cells, all from the same liquid sample.

17. The method of claim 14 , wherein the rare cells are white blood cells and the sample is enriched in white blood cells.

18. The method of claim 14 , wherein the rare cells are platelets and the sample is enriched in platelets.

19. The method of claim 14 , wherein the rare cells are red blood cells and the sample is enriched in red blood cells.

20. The method of claim 7 , wherein the liquid sample comprises a bodily fluid or an environmental liquid.

21. The method of claim 20 , wherein the bodily fluid comprises sweat, tears, ear flow, sputum, lymph, bone marrow suspension, cerebrospinal fluid, urine, saliva, semen, vaginal flow, cerebrospinal fluid, brain fluid, ascites, milk, secretions of the respiratory, intestinal or genitourinary tract, liquefied tissue sample, or amniotic fluid.

22. The method of claim 20 , wherein the rare cells are fetal cells and the liquid sample comprises maternal blood.

23. The method of claim 20 , wherein the rare cells are circulating cancer cells and the liquid sample comprises blood.

24. The method of claim 7 , wherein the particles of the specific type comprise cellular components, viruses, nucleic acids, lipids, proteins, protein complexes, or carbohydrates.

25. The method of claim 7 , wherein the array of obstacles comprises at least a first row and a second row, wherein the second row is displaced laterally relative to the first row so that particles in the liquid sample passing through gaps between obstacles in the first row are divided unequally to pass through two gaps between obstacles in the second row.

26. The method of clai

Assignees

• Massachusetts Gen Hospital
• Gpb Scientific Llc

Inventors

• Huang Lotien Richard
• Barber Thomas A
• Carvalho Bruce L
• Kapur Ravi
• Vernucci Paul
• Toner Mehmet
• Wang Zihua

Classifications

• G01N1/4077

  by other techniques involving separation of suspended solids · CPC title

• B01L3/502753

  characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation · CPC title

• B01L2400/0406

  capillary forces · CPC title

• B01L2400/0472

  Diffusion · CPC title

• B01L3/502761

  specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules · CPC title