Apparatus for cryopreparation, dry stabilization and rehydration of biological suspensions

A rapid cryofixation device, comprising:

• (a) a hollow nebulizer column having a closed top end and an open bottom end, the top end of said nebulizer column being connected to tubing through which a dry gas under positive pressure is provided so as to generate a flow of dry gas from the top end to the bottom end of said nebulizer column;
• (b) a nebulizer having a discharge nozzle, said nebulizer being positioned within said hollow nebulizer column in such a manner so that the discharge nozzle is directed to the open bottom end of the nebulizer column and so as to generate at least one laminar passage way through which a laminar flow of dry gas is generated and wherein said nebulizer is connected to a conduit through which a cryosolution, said cryosolution comprising a microscopic biological suspension, is supplied to the discharge nozzle so as to form a plurality of microdroplets of the cryosolution within the laminar flow of dry gas;
• (c) a cryogenic surface comprising a rotatably mounted hollow cylinder having an inner surface and an outer surface and a high thermal conductivity, the outer surface of said cylinder having a mirror finish and the inner surface of said cylinder being cooled by a liquid cryogen introduced into said cylinder by suitable connecting means, said cryogenic surface being positioned at the open bottom end of said nebulizer column so that the plurality of microdroplets within the laminar flow of dry gas encounter the cryogenic surface and are rapidly cooled to form a plurality of frozen microdroplets on said cryogenic surface;
• (d) means for rotating the hollow cylinder of the cryogenic surface;
• (e) a collector comprising a blade having an edge and means for holding the edge of said blade in a position so as to remove the plurality of frozen microdroplets from said cryogenic surface to form a frozen sample of said cryogenic solution;
• and
• (f) a removable sample vial holder, said holder having a plurality of openings in which sample vials are held, at least one of said openings being located so as to place the sample vial held therein in a position for collecting the frozen sample.

A rapid cryofixation device, comprising:

• (a) a hollow nebulizer column having a closed top end and a open bottom end, the top end of said nebulizer column being connected to tubing through which a dry gas under positive pressure is provided so as to generate a flow of dry gas from the top end to the bottom end of said nebulizer column;
• (b) an ultrasonic nebulizer having a discharge nozzle, said nebulizer being positioned within said hollow nebulizer column in such a manner so that the discharge nozzle is directed to the open bottom end of the nebulizer column and so as to generate at least one passage way through which a laminar flow of dry gas is generated and wherein said nebulizer is operated at a frequency between 25 and 60 kHz and is connected to a conduit through which a cryosolution, said cryosolution comprising a microscopic biological suspension, is supplied to the discharge nozzle so as to form a plurality of microdroplets of the cryosolution within the laminar flow of dry gas;
• (c) a cryogenic surface, said cryogenic surface being positioned at the open bottom end of said nebulizer column so that the plurality of microdroplets within the laminar flow of dry gas encounter the cryogenic surface and are rapidly cooled to form a plurality of frozen microdroplets on said cryogenic surface;
• (d) a collector comprising a blade having an edge and means for holding the edge of said blade in a position so as to remove the plurality of frozen microdroplets from said cryogenic surface to form a frozen sample of said cryogenic solution;
• and
• (f) means for collecting said frozen sample of said cryogenic solution.