COLLECTION OF SUSPENDED CELLS USING A TRANSFERABLE MEMBRANE

• US 20170268967A1
• Filed: 03/16/2016
• Published: 09/21/2017
• Est. Priority Date: 03/16/2016
• Status: Active Grant

• Current Assignee: Cellmax Ltd.
• Original Assignee: Cellmax Ltd.
• Inventors: Shao, Hung-Jen
• Granted Patent: US 10,107,726 B2
• Time in Patent Office: Days
• CPC Class Codes: B01D 63/081 Manufacturing thereof
• CPC Class Codes: B01D 63/088 Microfluidic devices comprising semi-permeable flat membranes
• CPC Class Codes: B01L 2300/048 enabling gas exchange, e.g. vents
• 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 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/126 Paper
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• 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 33/14 with filters, sieves or membranes
• CPC Class Codes: C12M 47/02 Separating microorganisms from the culture medium; Concentration of biomass separating microorganisms from their culture media C12N1/02
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 1/312 for samples mounted on planar substrates

Abstract

Devices, methods, and kits directed towards collecting and preparing cells using a separable sample collection layer may be configured to collect or treat cells from a liquid sample with mechanisms for easy transfer of the cells prior to analysis or imaging. The separable sample collection layer may comprise a porous membrane that cells may be collected on, and one or more support layers comprising tape with one or more adhesive coatings and release liner. The devices, methods and kits may be configured with support layers comprising cutouts that form vertically or horizontally oriented microchannels for efficiently removing undesirable liquid. Following collection and/or treatment, cells collected onto the porous membrane may be adhered to another surface for further processing or analysis.

Claims

1. A device for collecting cells of interest from a sample comprising:

a separable sample collection layer comprising a porous membrane, wherein the porous membrane is configured to receive the sample and collect the cells of interest;

a support layer coupled to, and in fluidic communication with the separable sample collection layer, wherein the support layer comprises an application zone configured to receive the sample, a channel fluidly coupled to the application zone, and a first outlet at a distal end of the channel, the first outlet defining a first flow path through which the sample flows out of the device; and

a solid support coupled to, and in fluidic communication with the support layer, wherein the solid support comprises a second outlet defining a second flow path through which the sample flows out of the device.

2. The device of claim 1, wherein the support layer comprises a release liner.

3. The device of claim 1, wherein the support layer comprises a low adhesion surface.

4. The device of claim 1, wherein a support layer comprises a double coated tape comprising a backing with adhesive and a release liner.

5. The device of claim 2, wherein the release liner comprises paper, plastic, polymer coated or any combination thereof

6. The device of claim 2, wherein a support layer comprises a polyethylene terephthalate (PET) film coated with acrylic adhesive, and a release liner.

7. The device of claim 1, wherein the solid support comprises plastic.

9. The device of claim 1, wherein the porous membrane comprises pore sizes up to 10 microns in diameter.

10. The device of claim 1, wherein the porous membrane comprises average porous membrane pore sizes that average 2-5 microns in diameter.

11. The device of claim 1, wherein the porous membrane comprises pore sizes that averages less than 2 microns in diameter.

12. The device of claim 1, wherein the porous membrane comprises polycarbonate.

13. The device of claim 1, wherein the porous membrane is circular and comprises a diameter equal to about 25 mm.

14. The device of claim 1, wherein the microchannel is circular and comprises a diameter equal to about 10 mm.

15. The device of claim 1, wherein the solid support and the support layer is in fluid communication through the porous membrane.

16. The device of claim 1, further comprising an absorbent pad coupled to, and in fluidic communication with the solid support.

17. The device of claim 1, further comprising one or more microchannels in fluid communication with the solid support and one or more of the support layers.

18. The device of claim 1, wherein the support layer further comprises one or more horizontal microchannels configured to prevent sagging of the porous membrane.

31. The device of claim 1, wherein the separable sample collection layer and the support layer are removably coupled to the solid support.

32. The device of claim 31, wherein the separable sample collection layer and the support layer are removably coupled to the solid support via an adhesive or release agent.

33. The device of claim 1, wherein the separable sample collection layer is parallel to, and located above the support layer, and wherein the support layer is parallel to, and located above the solid support.

34. The device of claim 1, further comprising a second support layer located adjacent to the separable collection layer.

35. The device of claim 34, wherein the second support layer is parallel to, and located above the separable sample collection layer, the separable sample collection layer is parallel to, and located above the support layer, and the support layer is parallel to, and located above the solid support.

36. The device of claim 1, wherein the second outlet is fluidly coupled to the channel.

37. The device of claim 36, wherein the second outlet is located beneath the channel.

38. The device of claim 1, wherein the first flow path and the second flow path are in opposite directions.

39. The device of claim 1, wherein the first flow path and the second flow path are substantially perpendicular to an orientation of the channel.

8. (canceled)

19. A method of collecting and preparing cells comprising:

a. applying biological sample to a separable sample collection layer comprising a support layer and a porous membrane, disposed against a solid support layer, wherein the cells are applied onto the porous membrane of the separable sample collection layer;

b. perfusing one or more liquid solutions through the porous membrane of a separable sample collection layer;

c. removing the separable sample collection layer from a supporting surface; and

transferring the separable sample collection layer to another surface for further processing or analysis.

20. The method of claim 19, wherein the other surface is a glass slide for imaging.

21. The method of claim 19, wherein the support layer is a piece of double coated tape with release liner.

22. (canceled)

23. (canceled)

24. (canceled)

25. (canceled)

26. (canceled)

27. (canceled)

28. (canceled)

29. (canceled)

30. (canceled)