Collection of suspended cells using a transferable membrane

• US 10,605,708 B2
• Filed: 09/06/2018
• Issued: 03/31/2020
• Est. Priority Date: 03/16/2016
• Status: Active Grant

• Current Assignee: Cellmax Ltd.
• Original Assignee: Cellmax Ltd.
• Inventors: Shao, Hung-Jen
• Primary Examiner(s): Nagpaul, Jyoti
• Application Number:
  US16/124,092
• Publication Number:
  US 20190242796A1
• Time in Patent Office: 572 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 method of collecting and preparing cells comprising:

(a) applying a biological sample comprising cells 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 the separable sample collection layer;

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

(d) transferring the separable sample collection layer to another surface.

2. The method of claim 1, wherein the another surface is a glass slide.

3. The method of claim 1, wherein the support layer comprises double coated tape.

4. The method of claim 3, wherein in a), the double coated tape is adhered to at least one of the porous membrane, a release liner, or the solid support layer.

5. The method of claim 1, wherein the removing the separable sample collection layer comprises separating the separable sample collection layer from the supporting surface.

6. The method of claim 1, wherein the method further comprises adhering the separable sample collection layer to the another surface.

7. The method of claim 1, wherein the transferring comprises transferring the seperable sample collection layer such that a sample collected by the separable sample collection layer is between the porous membrane and the another surface.

8. The method of claim 1, wherein prior to perfusing, the one or more liquid solutions comprise a foam, a lipid, or both.

9. The method of claim 1, wherein prior to perfusing, the one or more liquid solutions comprise circulating tumor cells (CTCs).

10. The method of claim 1, wherein the perfused liquid comprises an antibody, a buffer, a cell stain, a cell staining reagent, a reagent for fixing cells, or a surfactant.

11. The method of claim 10, wherein the perfused liquid interacts with cells of the biological sample such that the cells are fixed, stained, washed, bound to antibodies, or a combination thereof.

12. The method of claim 1, wherein the method further comprises processing or analyzing a sample collected on the separable sample collection layer after transferring the separable sample collection layer to the another surface.

13. The method of claim 12, wherein the processing or analyzing comprises imaging.

14. The method of claim 1, wherein the method further comprises directing a liquid using one or more microchannels or cutouts, wherein the liquid comprises at least one of the perfused liquid solutions or a liquid fraction of the biological sample.

15. The method of claim 14, wherein the directing occurs horizontally within layers.

16. The method of claim 14, wherein the directing occurs vertically between layers.

17. The method of claim 14, wherein the directing is assisted using gravity.

18. The method of claim 14, wherein the directing is assisted using pressure or a vacuum.

19. The method of claim 1, further comprising prior to c), removing a liquid fraction of the biological sample.

20. The method of claim 1, wherein in c) the separable sample collection layer comprises one or more cells of the cells that are applied onto the porous membrane of the separable sample collection layer.

21. The method of claim 1, wherein the support layer comprises an adhesive surface and a release liner.

22. The method of claim 21, wherein in a), the adhesive surface is adhered to the release liner.

23. The method of claim 22, wherein the removing comprises separating the adhesive surface from the release liner.

24. The method of claim 22, wherein the method further comprises adhering the adhesive surface to the another surface.

25. The method of claim 1, wherein the supporting surface comprises release paper.

26. The method of claim 25, wherein the separable sample collection layer comprises an adhesive adhered to the release paper, and the removing comprises separating the adhesive from the release paper.

27. The method of claim 1, wherein removing comprises peeling the separable sample collection layer from the supporting surface using mechanical force.

28. The method of claim 27, wherein the removing is performed such that cells in the biological sample remain viable.

29. The method of claim 1, wherein the method further comprises fixing one or more cells of the biological sample with paraformaldehyde.