A differential staining technique for simultaneous visualization of mitotic spindle and chromosomes in mammalian cells

Wissinger, W.L.; Estervig, D.N.; Wang, R.J.

Stain Technology 56(4): 221-226

1981


ISSN/ISBN: 0038-9153
PMID: 6171054
DOI: 10.3109/10520298109067315
Accession: 004571010

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Abstract
A new technique was devised for staining the mitotic spindle in mammalian cells while preserving spindle structure and chromosome number. The cells are trypsinized and fixed with a 3:1 methanol:acetic acid solution containing 4 mM MgCl2 and 1.5 mM CaCl2 at room temperature. The cells are placed on slides and treated with 5% perchloric acid before staining with a 10% acetic acid solution containing safranin O and brilliant blue R. The preserved spindles appear dark blue against a light cytoplasmic background with chromosomes stained bright red. Individual chromosomes and chromatids are clearly visible. Positioning of the chromosomes relative to the spindle apparatus is readily ascertained, allowing easy study of mitotic spindle and chromosome behavior.