Crystal Violet Staining,细胞结晶紫染色
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Iruela-Arispe Lab
University of California, Los Angeles Crystal Violet (CV) Staining of Cells and Clone counting
Timothy Lane
Stocks.
3.7% Paraformaldehyde (PFA) or 10% Formalin
0.05% Crystal Violet in Distilled Water (Filter at 0.45um before use)
Fix the cells for 5 min. with 3.7% PFA
STAIN them 30 minutes with 0.05%CV
Wash 2x with tap water,
Drain them inverted for a couple minutes.
The dishes can then be photographed or scanned for colony counts
READING CV STAIN (OD 540)
add a third to half the total well volume of straight methanol to
solubilize s the dye,
Read the plates (OD 540) either directly or by taking an aliquot to another
plate.
The cells never lyse and morphology is extremely well preserved by the
combined formalin/methanol fixations.
ALTERNATIVE PROTOCOL:
USE 0.1 % CV in 2 % ethanol in 0.1 M borate ph9
NO NEED TO PRE-FIX