Integrated microfluidic platform for selective extraction of single-cell mRNA

Patent metadata

Field	Value
Publication number	US-10549277-B2
Application number	US-201715697898-A
Country	US
Kind code	B2
Filing date	Sep 7, 2017
Priority date	Oct 14, 2015
Publication date	Feb 4, 2020
Grant date	Feb 4, 2020

Abstract

Disclosed is a lab-on-a-chip platform comprising a microfluidic device sealed with a thin membrane. An analysis probe configured to penetrate the thin membrane to access a sample trapped in the microfluidic device.

First claim

What is claimed is:

1. A lab-on-a-chip platform comprising:
a microfluidic device comprising:
an analysis region comprising a single cell trapping array; and
a membrane sealing the analysis region from an ambient environment; and
an external dielectric nanotweezer (DENT)

wherein the membrane is configured to be punctured by the external DENT to form a hole therein to allow the external DENT to access the microfluidic sample.

2. The lab-on-a-chip platform of claim 1 , wherein the external DENT comprises an atomic force microscope probe.

3. The lab-on-a-chip platform of claim 2 , wherein the external DENT is configured to extract intracellular components from a trapped sample comprising an array of single cells in one or more microfluidic channels.

4. The lab-on-a-chip platform of claim 3 , wherein the external DENT is configured to analyze mRNA expression levels of the extracted intracellular components.

5. The lab-on-a-chip platform of claim 1 , wherein the external DENT comprises a micro-injector.

6. The lab-on-a-chip platform of claim 1 , wherein the microfluidic device further comprises a microfluidic sample processing region upstream from the analysis region.

7. The lab-on-a-chip platform of claim 6 , wherein the sample processing regions include sample sorting or sample separation region.

8. The lab-on-a-chip platform of claim 1 , wherein the membrane is configured to be resealable such that after the hole is formed therein by the micro-manipulating instrument, the hole seals after removal of the micro-manipulating instrument.

9. The lab-on-a-chip platform of claim 1 , wherein the membrane has a thickness of less than 5 μm.

10. The lab-on-a-chip platform of claim 1 , wherein the membrane comprises an elastomer.

11. The lab-on-a-chip platform of claim 10 , wherein the elastomer comprises PDMS.

12. A method of manufacturing a lab-on-a-chip platform according to claim 1 , the method comprising:
providing a microfluidic device comprising a microfluidic channel and a membrane configured to seal the microfluidic device from an ambient environment; and
bonding the sealed microfluidic device on a substrate;
wherein the membrane has a thickness less than 5 μm, and wherein the membrane encloses an analysis region of the microfluidic channel.

13. The method of claim 12 , wherein the membrane comprises an elastomer.

14. The method of claim 13 , wherein the elastomer comprises PDMS.

15. The method of claim 12 , wherein at least one of an upstream sample flow region and a downstream sample flow region of the microfluidic channel is enclosed by the membrane.

16. The method of claim 12 , wherein at least one of a sample processing region and a post-processing region of the microfluidic channel is enclosed by a structure that is thicker than the membrane.

17. A method of analyzing a sample, comprising:
providing a lab-on-a-chip platform according to claim 1 ;
disposing a sample containing cells on a portion of the analysis region for isolating the cells,
disposing the microfluidic component on, under, beside or in an external DENT having a probe with a tip;
puncturing the membrane with the tip of the probe of the external micro-manipulating instrument; and
manipulating an individual cell of the sample in the microfluidic channel using the tip of the probe.

18. The method of claim 17 , wherein the membrane comprises an elastomer.

19. The method of claim 18 , wherein the elastomer comprises PDMS.

20. The method of claim 17 , wherein the external DENT comprises an atomic force microscope.

21. The method of claim 17 , wherein the analysis region comprises a single cell trapping array for trapping a plurality of individual cells.

Assignees

• Univ California

Inventors

• Lee Abraham P
• Wickramasinghe H Kumar
• Lee Do-Hyun
• Tao Yinglei
• Li Xuan

Classifications

• C12N15/101

  by chromatography, e.g. electrophoresis, ion-exchange, reverse phase · CPC title

• B01L2200/0668

  Trapping microscopic beads · CPC title

• B01L3/502715

  characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces · CPC title

• B01L2200/0652

  Sorting or classification of particles or molecules · CPC title

• C12Q1/6806

  Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay (C12Q1/6804 takes precedence) · CPC title