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Slow‐speed freezing of chemically unfixed biological tissues and long‐term storage of frozen samples for cyroscanning electron microscopy
Author(s) -
Adler Klaus,
Kruse Joachim,
Kunze Gotthard
Publication year - 1996
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/(sici)1097-0029(19960215)33:3<262::aid-jemt3>3.0.co;2-s
Subject(s) - liquid nitrogen , allium , agar , electron microscope , yeast , agar plate , biology , scanning electron microscope , botany , materials science , biophysics , chemistry , optics , biochemistry , composite material , bacteria , genetics , physics , organic chemistry
We describe a procedure in which plant tissue as well as a yeast culture on agar are frozen with slow cooling rates for observation of surface structures in a cryoscanning electron microscope. A system is also presented for long‐term storage of frozen specimens under liquid nitrogen, in which the material is maintained for direct observation. Some small tools are described, which are essential for making preparations using slow‐speed freezing and for the storage of prepared samples. Three examples of preparations with different complications are given: the “sculptures” on the surface of a leaf of Allium schoenoprasum, an early stage of flower development of Allium cernuum, and a part of an agar‐grown colony of Arxula adeninivorans. In our experience, it is possible to store fully hydrated samples under the described conditions for more than a year without damaging the fine structures. © 1996 Wiley‐Liss, Inc.