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Fixation of the cytoskeleton
If components of the cytoskeleton are of interest, cells can be extracted prior to fixation.
- Rinse cells with PBS.
- Incubate cells in extraction buffer (100 mM Pipes-KOH, 2 mM EGTA, 1 mM MgCl2, 0.5% Triton X-100, pH 6.8) for 1 to 2 minutes at room temperature.
- Fix in methanol or formaldehyde/glutaraldehyde (see above).
- Rinse cells with PBS.