Methods and apparatuses for sorting objects in forensic DNA analysis and medical diagnostics

• US 20090075826A1
• Filed: 09/11/2008
• Published: 03/19/2009
• Est. Priority Date: 09/13/2007
• Status: Active Grant

• Current Assignee: Arryx, Inc. (Haemonetics Corporation)
• Original Assignee: Arryx, Inc. (Haemonetics Corporation)
• Inventors: Chakrabarty, Tania
• Granted Patent: US 7,998,676 B2
• Time in Patent Office: Days
• US Class Current: 506/6
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2300/0681 Filter
• 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 2400/0454 radiation pressure, optical tweezers
• 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 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: G01N 2001/007 Devices specially adapted for forensic samples, e.g. tamper-proofing, sample tracking
• CPC Class Codes: G01N 35/00029 provided with flat sample substrates, e.g. slides G01N35/028 takes precedence

Abstract

The present invention relates to an apparatus and method of sorting objects and identifying the objects in a forensics sample, including using holographic optical trapping to sort objects from contaminants, and performing (single cell) PCR-based STR analysis on the objects to determine their identification. In addition, the chip used as a support for sorting the objects can also be used for performing single cell PCR-based STR analysis. In another embodiment, a microfluidics chip is used to stream the sample and sort the objects, before single cell PCR-based STR analysis is performed. The chip used for sorting utilizing HOT in the absence or presence of microfluidic streaming and sorting can also be the same as that used for the single cell PCR-based STR analysis.

Claims

1. A method of sorting objects and identifying the objects in a sample, comprising:

providing a holographic optical trapping apparatus;

providing a sample, said sample containing objects to be separated;

optically trapping said objects in said sample;

sorting said objects using said optical trapping into wanted and unwanted objects;

removing at least one wanted object from said sample; and

performing PCR-based STR analysis on said wanted object to determine an identification of said wanted object.

2. The method according to claim 1, wherein prior to said optical trapping step, said method further comprises:

providing said sample in a fluidic stream in a microfluidics chip; and

streaming said sample through said microfluidics chip.

3. The method according to claim 2, wherein each of the wanted objects are sorted into individual chambers.

4. The method according to claim 2, wherein said sorting step and said performing step are performed on a single microfluidics chip.

5. The method according to claim 1, wherein the wanted objects are cells and the unwanted objects are contaminants.

6. The method according to claim 1, wherein the wanted objects are cancer cells and the unwanted objects are healthy cells, or vice versa.

7. The method according to claim 3, wherein the performing step is single cell PCR-based STR analysis.

8. The method according to claim 5, wherein the microfluidics chamber includes an input chamber which contains said sample, and said sample is streamed from said input chamber into at least one output chamber.

9. The method according to claim 8, further comprising:

providing a filter at an outlet of said input chamber.

10. The method according to claim 9, further comprising:

providing valves in said microfluidics chamber to prevent backflow.

11. The method according to claim 1, wherein said sorting and PCR performing steps are automated and multiplexed.

12. A method of sorting objects and identifying the objects in a sample, comprising:

providing a microfluidics chip having an input chamber and at least one output chamber;

providing a sample in said input chamber, said sample containing objects to be sorted;

sorting said objects using a fluid stream in said input chamber, such that said objects are sorted into wanted and unwanted objects;

removing said wanted objects into at least one said output chamber; and

performing PCR-based STR analysis on each said wanted object to determine an identification of said wanted object.

13. The method according to claim 12, wherein said sorting step and said performing step are performed on a single microfluidics chip.

14. The method according to claim 12, wherein the wanted objects are cells and the unwanted objects are contaminants.

15. The method according to claim 12, wherein the performing step is single cell PCR-based STR analysis.

16. The method according to claim 12, wherein the wanted objects are normal cells and the unwanted objects are cancerous cells, or vice versa.

17. The method according to claim 12, further comprising:

providing a filter at an outlet of said input chamber.

18. The method according to claim 17, further comprising:

providing valves in said microfluidics chamber to prevent backflow.

19. The method according to claim 12, wherein said sorting and PCR performing steps are automated and multiplexed.

20. An apparatus for sorting objects, comprising:

a holographic optical trapping apparatus containing a support on which a sample is disposed, said optical trapping apparatus which optically traps and sorts wanted objects from unwanted objects in said sample; and

an apparatus for performing PCR-based STR analysis on said wanted objects.

21. The apparatus of claim 20, wherein said wanted objects are cells and said unwanted objects are contaminants.

22. The apparatus of claim 20, wherein said wanted objects are normal cells and said unwanted objects are cancerous cells or vice versa.

23. The apparatus of claim 20, wherein said PCR apparatus performs single cell PCR-based STR analysis.

24. The apparatus of claim 20, further comprising:

a microfluidics chip for streaming said sample prior to optical trapping of said wanted objects.

25. The apparatus of claim 20, wherein said support and said apparatus for performing said PCR-based STR analysis are a single chip.

26. The apparatus of claim 20, further comprising:

a plurality of valves disposed in said microfluidics chamber, to prevent backflow.

27. The apparatus according to claim 20, further comprising:

a filter disposed at an outlet of said inlet chamber.

28. The apparatus according to claim 20, wherein said sorting and PCR performing steps are automated and multiplexed.

29. An apparatus for sorting objects, comprising:

a microfluidics chip containing an input chamber and at least one output chamber, said input chamber containing a sample;

wherein said microfluidics chip streams said sample such that wanted objects are sorted from unwanted objects from said input chamber to said at least one output chamber; and

an apparatus for performing PCR-based STR analysis on said wanted objects.

30. The apparatus of claim 29, further comprising:

a holographic optical trapping apparatus for sorting said wanted objects from said unwanted objects from said input chamber into said at least one output chamber.

31. The apparatus of claim 29, wherein said microfluidics chip and said apparatus for performing said PCR-based STR analysis are a single chip.

32. The apparatus of claim 29, wherein said objects are cells, and said unwanted objects are contaminants.

33. The apparatus of claim 29, wherein said wanted objects are normal cells and said unwanted objects are cancerous cells.

34. The apparatus of claim 29, further comprising:

a plurality of valves disposed in said microfluidics chamber, to prevent backflow.

35. The apparatus according to claim 29, further comprising:

a filter disposed at an outlet of said inlet chamber.

36. The apparatus according to claim 29, wherein said sorting and PCR performing steps are automated and multiplexed.