Filter support with a phase-changing medium
The present invention relates to a method for processing isolated cells, in particular rare cells such as circulating tumor cells, to render them suitable for optical imaging, comprising the steps of (a) depositing filter material comprising captured cells on a phase-change medium; (b) melting said phase-change medium until the medium is spread out below said filter material; and (c) lowering the temperature of said phase-change medium until the medium solidifies, resulting in an optically flat filter comprising said cells at a fixed position. The method may additionally comprise an initial step of capturing a cell on a filter material. The phase-change medium, preferably paraffin wax, may comprise a porous, or mesh-like structure allowing the passage of a fluid through the medium. The phase-change medium may further be mounted on a carrier such as a glass carrier or a polymer material carrier. The present invention further relates to processed filter material comprising isolated cells suitable for optical imaging, which is obtainable by the method; the use of such filter material for diagnostic, histological, microbiological, biochemical, oncologic, or hematologic analysis, as well as a device for isolating cells or processing cells to render them suitable for optical imaging.
1. Method for processing isolated cells to render them suitable for optical imaging, comprising the following steps:
(a) depositing filter material comprising captured cells on a phase-change medium, said phase-change medium being mounted on a carrier;
(b) melting said phase-change medium until the medium is spread out below said filter material; and
(c) lowering the temperature of said phase-change medium until the medium Solidifies, resulting in a filter comprising said cells in a single optical plane and at a fixed position.
2. The method of claim 1 , additionally comprising the initial step of capturing a cell on a filter material.
3. The method of claim 1 , wherein said filter material is capable of size excluding cells to be isolated from a fluid or solution having a pore size of about 0.5 to 20 μm and wherein said filter material is a polymer filter.
4. The method of claim 1 , wherein said phase-change medium is a medium which melts and solidifies in a temperature range of about 25° C to 70° C.
5. The method of claim 1 , wherein said phase-change medium comprises a porous, mesh-like, open, or partially open structure and/or comprises channel, passage or pipe elements, wherein said structure or elements allow the passage of a fluid through said medium.
6. The method of claim 4 , wherein said medium is a hydro wax.
7. The method of claim 1 , wherein said carrier is a glass carrier or a polymer material carrier, and wherein said carrier comprises a position marker.
8. The method of claim 1 , wherein said method comprises as additional step an activity selected from the group of staining, fixating, releasing, mechanically manipulating, biologically manipulating and biochemically manipulating said isolated cell(s).
9. The method of claim 8 , wherein said activity comprises culturing said isolated cell(s) via flowing a nutrient fluid through a porous, meshlike, open, or partially open phase-change medium.
10. The method of claim 1 , wherein said cell is a circulating tumor cell, or a bacterial cell.
11. The method of claim 1 , wherein said optical imaging is bright field microscopy, dark field microscopy, phase contrast microscopy, differential interference contrast microcopy, fluorescence microscopy, atomic force microscopy, confocal laser scanning microscopy or super resolution microscopy.
12. A method for optical imaging for diagnostic, histological, microbiological, biochemical, oncologic, or hematologic analysis comprising the step of imaging a processed filter material comprising isolated cells which is obtained by the method according to claim 1 .
13. Device for processing cells to render them suitable for optical imaging, comprising:
(a) a filter material configured to capture a cell;
(b) a removable pressure delivery system coupled to the filter material configured to isolate cells onto the filter material;
(c) a layer of phase-change medium;
(d) a means for transporting said filter material and/or said layer of phase-change medium to a carrier;
(e) a heating unit configured to melt said phase-change medium until the medium is spread out below said filter materials; and
(f) a cooling unit configured to lower the temperature of said phase-change medium until the medium solidifies and provides a filter comprising said cells in a single optical plane and at a fixed position.