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Retention of vacuole contents of plant cells during fixation
Author(s) -
DONG Z.,
McCULLY M. E.,
CANNY M. J.
Publication year - 1994
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.1365-2818.1994.tb03488.x
Subject(s) - vacuole , glutaraldehyde , fixation (population genetics) , biophysics , permeability (electromagnetism) , plant cell , chemistry , cytoplasm , formaldehyde , microbiology and biotechnology , phosphate buffered saline , biology , biochemistry , chromatography , membrane , gene
Summary Changes in the semi‐permeability of tonoplast during fixation were studied using beetroot tissue. Using vacuole betacyanin as the indicator, the permeability of the tonoplast was assessed by the leakage of this pigment as determined by changes in the optical density of the solution bathing the tissue. Cryo‐analytical scanning electron microscopy was used to monitor the changes in ion concentration in the cells during fixation. Fixatives were 3% glutaraldehyde or 4% formaldehyde in 0·025 m phosphate buffer at room temperature or on ice. Results showed that glutaraldehyde, especially at low temperature, takes as long as 30 h to disrupt the semi‐permeability of the tonoplast of beetroot cells, while in formaldehyde on ice, these cells begin to lose their selective permeability in 15 min. This study confirms that the semi‐permeability of the tonoplast may not be lost until long after the cytoplasm has been fixed and suggests that this explains why cold fixation in 3% glutaraldehyde for about 12 h has become the most reliable standard procedure for the successful preservation of vacuolated plant cells.