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Fixation induces differential tip morphology and immunolocalization of the cytoskeleton in pollen tubes
Author(s) -
He Yi,
Wetzstein Hazel Y.
Publication year - 1995
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1995.tb05128.x
Subject(s) - fixative , egta , fixation (population genetics) , biology , cytoskeleton , botany , microfilament , pollen , biophysics , anatomy , calcium , cytoplasm , microbiology and biotechnology , chemistry , biochemistry , cell , organic chemistry , gene
The effects of chemical fixation on tip morphology and immunolocalization of the cytoskeleton in Tradescantia virginiana pollen tubes were evaluated using lour different fixation protocols differing in fixative type/concentration, fixation time, buffer system and additives. Apical regions were much more sensitive to fixation manipulations than more basipetal areas. The presence of the calcium chelator EGTA at 5 mM led to tip rupture and/or swelling in over 80% of germinated grains. However, low or no EGTA levels during fixation resulted in poor immunolocalizations, although lips had more normal morphology. Double fixation in which pollen tubes were first treated for a short period with higher fixative and lower EGTA (0.5 mM) concentrations, followed by lower fixative and higher EGTA (5.0 mM) concentrations, resulted in both improved preservation of pollen lube tip morphology and microfilament/microtubule localizations.