madrid2i

Resumen

The present invention relates to a method and system for detecting, manipulating and isolating circulating tumour cells found in organic liquids by a simplified laser-assisted transfer process. A liquid sample of the organic liquid that contains an enriched population of tumour cells and a population of non-tumour cells is extended in a donor substrate with an intermediate layer that comprises a semi-transparent polymeric adhesive tape. Fluorescent staining of at least one of the cell populations found in the liquid sample is required in order to identify by optical microscopy and fluorescence the location of the cells to be manipulated and/or isolated. The energy from the laser beam is absorbed by the polymeric adhesive tape, which results in the formation of a blister that interacts mechanically with the liquid sample and said interaction can induce the ejection of a portion of the liquid sample that comprises the selected tumour cells or non-tumour cells, which can be a single cell or a group of cells, towards the receiving substrate.

Reivindicaciones

1. A method for detecting and manipulating tumor cells found in organic fluids comprising the steps of: a) providing a liquid specimen 400 of the organic fluid containing an enriched population of tumor cells and a population of non-tumor cells; b) fluorescently staining at least one of the two cell populations wherein a different fluorescence dye is used for each population when both populations are stained; c) providing a donor substrate 300 comprising a transparent support 100 and a semi-transparent polymeric adhesive tape 200 adhered to the transparent support 100; d) dispensing and spreading the liquid specimen 400 onto the semi-transparent polymeric adhesive tape 200; e) providing a receiving substrate 500; f) placing the receiving substrate 500 facing the surface of the donor substrate 300 where the liquid specimen 400 is spread maintaining a separation distance between them; g) visualizing the same portion of the liquid specimen 400 by means of optical and fluorescence microscopy 700; h) comparing the optically and fluorescently obtained images to identify the location of tumor cells or non-tumor cells; i) providing a laser beam 810 focused on the interface between the transparent support 100 and the semi-transparent polymeric adhesive tape 200 of the donor substrate 300; j) directing the laser beam 810 to the identified location of at least one tumor cell or at least one non-tumor cell; so that the laser beam 810 energy is absorbed by the semi-transparent polymeric adhesive tape 200 at the identified location; giving place to the formation of a blister on the polymeric adhesive tape 200 that generates a mechanical perturbation in the liquid specimen leading to the movement of at least one cell or cluster of cells. 2. The method according to claim 1, wherein the blister mechanically propels a portion of the liquid specimen towards the receiving substrate 500, the propelled liquid comprising at least one tumor cell without any non-tumor cells. 3. The method according to claim 1, wherein the blister mechanically propels a portion of the liquid specimen towards the receiving substrate 500, the propelled liquid comprising at least one non-tumor cell without any tumor cells. 4. The method according to claims 2 or 3, wherein the portion of the liquid specimen propelled by the laser beam comprises a single cell. 5. The method according to claims 2 or 3, wherein the portion of the liquid specimen propelled by the laser beam comprises a cluster of cells. 6. The method according to any of the claims 2 to 5, wherein only the tumor cells or only the non-tumor cells are stained. 7. The method according to claim 6, wherein the portion of the liquid specimen propelled by the laser beam comprises at least one non-stained cell. 8. The method according to claim 6, wherein the portion of the liquid specimen propelled by the laser beam comprises at least one stained cell. 9. The method according to claim 1, wherein the generated blister leads to the disaggregation of a cluster of cells. 10. The method according to any preceding claim, wherein tumor cells comprise at least one of circulating tumor cells and disseminated tumor cells. 11. The method according to any preceding claim, wherein the non-tumor cells comprise at least one of hematopoietic cells and white blood cells. 12. A system for detecting, manipulating and isolating tumor cells from an organic fluid comprising: a donor substrate comprising a transparent support and a semi-transparent polymeric adhesive tape adapted to be adhered on one side of the transparent support and further adapted to receive, on the side opposite to the transparent support, a liquid sample of the organic fluid containing an enriched population of tumor cells and a population of non-tumor cells wherein at least one of the two cell populations are fluorescently stained, a receiving substrate arranged at a distance from the donor substrate while facing the liquid sample; a visualization system adapted to provide a visualization of the same portion of the liquid sample by means of optical and fluorescence microscopy and further adapted to compare the optically and fluorescently obtained images, a laser system adapted to focus a laser beam on the interface between the transparent support and the semi-transparent polymeric adhesive tape of the donor substrate and further adapted to direct the laser beam to the location of at least one tumor cell or at least one non-tumor cell so that the laser beam energy is absorbed by the semi-transparent polymeric adhesive tape giving place to the formation of a blister on the polymeric adhesive tape that generates a mechanical perturbation in the liquid specimen leading to the movement of at least one cell or cluster of cells. 13. A system for detecting, manipulating and isolating tumor cells from an organic fluid according to claim 12, wherein the semi-transparent polymeric adhesive tape comprises at least one thermoplastic polyimide sheet. 14. A system for detecting, manipulating and isolating tumor cells from an organic fluid according to claims 12 or 13 wherein the semi-transparent polymeric adhesive tape comprises at least one layer of a transparent adhesive silicone.

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