Electrophoretic separation devices and methods for using the same

Patent metadata

Field	Value
Publication number	US-10267795-B2
Application number	US-201414650520-A
Country	US
Kind code	B2
Filing date	Mar 6, 2014
Priority date	Mar 7, 2013
Publication date	Apr 23, 2019
Grant date	Apr 23, 2019

Abstract

Electrophoretic separation devices and methods for using the same are provided. Aspects of the devices include a polymeric separation medium that includes a plurality of microwells. Also provided are methods, systems and kits in which the subject devices find use. The devices and methods find use in a variety of different electrophoretic separation applications.

First claim

That which is claimed is:

1. A device comprising:
a polymeric separation medium comprising a plurality of microwells, wherein the polymeric separation medium comprises electromagnetic radiation-activatable functional groups that covalently bond to one or more sample components of interest in the separation medium upon application of electromagnetic radiation, wherein the electromagnetic radiation-activatable functional groups are co-polymerized with the polymeric separation medium, and wherein the microwells have a width of 100 μm or less and a depth of 100 μm or less.

2. The device of claim 1 , further comprising a solid support contacting a surface of the polymeric separation medium, wherein the device comprises at least one channel through a portion of one or more of the polymeric separation medium and the solid support.

3. The device of claim 1 , wherein the microwells are arranged as an array of microwells in the polymeric separation medium.

4. The device of claim 3 , wherein the microwells comprise an open end on the surface of the polymeric separation medium and an opposing closed end in the polymeric separation medium.

5. The device of claim 4 , wherein the open end of the microwell has a width greater than the closed end of the microwell.

6. The device of claim 1 , wherein the polymeric separation medium comprises a central well comprising a plurality of microwells positioned on the periphery and in fluid communication with the central well.

7. The device of claim 6 , wherein each microwell comprises an open end in fluid communication with the central well and an opposing closed end in the polymeric separation medium.

8. The device of claim 6 , wherein the microwells are arranged around substantially the entire periphery of the central well.

9. The device of claim 1 , wherein the polymeric separation medium comprises 100 or more microwells.

10. A method comprising:
contacting a sample with a polymeric separation medium of claim 1 ;
applying an electric field to the polymeric separation medium in a manner sufficient to move at least some components of the sample from the microwell into the polymeric separation medium to produce separated sample components in the polymeric separation medium; and
immobilizing the separated sample components in the polymeric separation medium by applying electromagnetic radiation to the electromagnetic radiation-activatable functional groups.

11. The method of claim 10 , wherein the sample comprises cells and/or cellular components.

12. The method of claim 11 , further comprising lysing the cells to produce the cellular components in the sample.

13. The method of claim 11 , further comprising incubating the cells to produce the cellular components in the sample.

14. The method of claim 10 , further comprising detecting the separated sample components.

15. The method of claim 14 , wherein the detecting comprises contacting the separated sample components with an analyte detection reagent.

16. The method of claim 15 , further comprising contacting the separated sample components with a second analyte detection reagent.

17. The method of claim 10 , further comprising imaging the polymeric separation medium to produce an image of the separated sample components.

18. The method of claim 10 , wherein the contacting the sample with the polymeric separation medium comprises positioning at least some components of the sample into one or more microwells.

19. The method of claim 18 , wherein at least some of the components in the sample are bound to magnetic beads, and the positioning comprises applying a magnetic force to the sample.

20. A kit comprising:
a device according to claim 1 ; and
a packaging containing the device.

21. The device of claim 1 , wherein each microwell accommodates a single cell.

22. The device of claim 1 , wherein each microwell holds a volume of about one nanoliter.

23. The device of claim 1 , wherein the device is configured for single cell analysis.

24. The device of claim 1 , wherein the electromagnetic radiation-activatable functional groups comprise benzophenone functional groups.

Assignees

• Univ California

Inventors

• Herr Amy E
• Hughes Alex James
• Sinkala Elly
• Duncombe Todd A
• Yamauchi Kevin A
• Vlassakis Julea
• Kang Chi-Chih
• Xu Zhuchen
• Lin Robert

Classifications

• G01N27/44704

  Details; Accessories · CPC title

• G01N27/44721

  by optical means · CPC title

• G01N33/561 Primary

  Immunoelectrophoresis · CPC title

• B01L3/5027

  by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip · CPC title

• B01L2300/0645

  Electrodes · CPC title