（交流）介绍6种针对免疫荧光染色的细胞固定方法
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1、Methanol-Acetone Fixation
Fix in cooled methanol, 10 min at –20 °C.
Remove excess methanol.
Permeabilize with cooled acetone for 1 minute at –20 °C.
2、Paraformaldehyde-Triton Fixation
Fix in 3-4% paraformaldehyde for 10-20 min.
Rinse briefly with PBS.
Permeabilize with 0.5% Triton X-100 for 2-10 min.
3、Paraformaldehyde-Methanol Fixation
Fix in 3-4% paraformaldehyde for 10-20 min.
Rinse briefly with PBS.
Permeabilize with cooled methanol for 5-10 min at –20 °C.
4、PEM-Ethanol Fixation
Fix in PEM buffer for 10 min.
Rinse twice, briefly, with PBS.
Permeabilize with cooled ethanol for 5-10 min at –20 °C.
PEM Buffer: 0.1M PIPES, 5 mM EGTA, 2mM MgCl2 .6H2O. Bring to pH 6.8, using NaOH solution .
5、Acetone - Triton X-100 Fixation
Rinse cells 2 times with HBSS at 37°C.
Fix cells by incubation in acetone at RT for 5 min.
Rehydrate cells for 10 min in PBS.
Permeabilize cells with in 0.2% Triton X-100 in PBS for 5 min.
Rinse samples 3 times, for one minute each time, in PBS.
6、Methanol - Acetone Fixation
Briefly rinse cells with PBS at 37°C.
Fix cells with -20°C acetone-methanol(1:1) for 10 min (do it in -20°C freezer).
Either: a) airdry cells and store, or
b) Rehydrate cells in PBS for 10 min at RT.
谢谢笑佛站友，还请能够规范发贴。
GOOD LUCK!
Fix in cooled methanol, 10 min at –20 °C.
Remove excess methanol.
Permeabilize with cooled acetone for 1 minute at –20 °C.
2、Paraformaldehyde-Triton Fixation
Fix in 3-4% paraformaldehyde for 10-20 min.
Rinse briefly with PBS.
Permeabilize with 0.5% Triton X-100 for 2-10 min.
3、Paraformaldehyde-Methanol Fixation
Fix in 3-4% paraformaldehyde for 10-20 min.
Rinse briefly with PBS.
Permeabilize with cooled methanol for 5-10 min at –20 °C.
4、PEM-Ethanol Fixation
Fix in PEM buffer for 10 min.
Rinse twice, briefly, with PBS.
Permeabilize with cooled ethanol for 5-10 min at –20 °C.
PEM Buffer: 0.1M PIPES, 5 mM EGTA, 2mM MgCl2 .6H2O. Bring to pH 6.8, using NaOH solution .
5、Acetone - Triton X-100 Fixation
Rinse cells 2 times with HBSS at 37°C.
Fix cells by incubation in acetone at RT for 5 min.
Rehydrate cells for 10 min in PBS.
Permeabilize cells with in 0.2% Triton X-100 in PBS for 5 min.
Rinse samples 3 times, for one minute each time, in PBS.
6、Methanol - Acetone Fixation
Briefly rinse cells with PBS at 37°C.
Fix cells with -20°C acetone-methanol(1:1) for 10 min (do it in -20°C freezer).
Either: a) airdry cells and store, or
b) Rehydrate cells in PBS for 10 min at RT.
谢谢笑佛站友，还请能够规范发贴。
GOOD LUCK!
