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Freezing in sealed capillaries for preparation of frozen hydrated sections
Author(s) -
YAKOVLEV S.,
DOWNING K.H.
Publication year - 2011
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2011.03530.x
Subject(s) - cryoprotectant , ice crystals , crystallization , freezing point , synchrotron , freeze drying , congelation , materials science , sample preparation , dextran , volume (thermodynamics) , chromatography , chemistry , cryopreservation , optics , thermodynamics , embryo , physics , organic chemistry , biology , microbiology and biotechnology
Summary We have investigated the freezing of specimens in a confined volume for preparation of vitreous samples for cryosectioning. With 15% dextran as a cryoprotectant, a sample sealed in a copper tube begins to freeze into crystalline ice when plunged into liquid ethane. Crystallization rapidly causes an increase in the pressure to the point that much of the sample freezes in a vitreous state. We used synchrotron X‐ray diffraction of samples frozen with various amounts of dextran to characterize the ice phases and crystal orientation, providing insights on the freezing process. We have characterized cryosections obtained from these samples to explore the optimum amount of cryoprotectant. Images of cryosectioned bacteria frozen with various levels of cryoprotectant illustrate effects of cryoprotectant concentration.