High-pressure freezing of cell suspensions in cellulose capillary tubes

A procedure for efficient cryoimmobilization of large volumes of cell suspensions or micro-organisms by high-pressure freezing is described. This procedure uses transparent, porous cellulose capillary tubes with an inner diameter of 200 microns, into which the suspensions are drawn by capillary action. The tubes are processed by high-pressure freezing and freeze-substitution as if they were tissue samples. Centrifugation of suspensions at low temperatures is no longer necessary and cryopreparation is greatly facilitated. A very high yield of adequately frozen specimens is obtained due to the constant, defined sample geometry. This approach can also be used to process suspensions by conventional chemical fixation, eliminating the need to embed pellets in low-melting-point agarose, for example, prior to chemical fixation. The preparation procedure is demonstrated with suspensions of nematodes, paramecia and bacteria.