Protocol according to “Romeis - Mikroskopische Technik” (German), 19th edition, ISBN-13: 978-3642551895.
The steps 2-4 may be also replaced by fixing in cold acetone (-20 °C) or in methanol (-20 °C) for 10-20 minutes each. My personal favorite is a 1:1 (v/v) mixture of ethanol and dimethyl ether (-20 °C, NOT more than 3-5 minutes!).
If there are problems with staining of membranbe-bound proteins, skip permeabilization and/or don't use lipophilic agents (like ether) for fixation of the cells.
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