LABELING OF CELLS WITH FLUORESCENT PHALLOIDIN

1. Wash 3 x 5 min PBS-Ca²⁺

2. Fix cells in 4% Formaldehyde in 0.1 M Phosphate buffer pH 7.4, 20 min, 22°C

3. Wash 3 x 5 min PBS-Ca²⁺

4. Permeabilization: 0.1% Triton X100 in PBS-Ca²⁺ 20 min, 22°C

5. Wash 3 x 5 min PBS-Ca²⁺

6. Incubation with Phalloidin-Rhodamine (Stock = 100 µg/ ml), dilution 1/100 in PBS with 0.1% BSA, 1 h, 4°C

7. Wash 5 x 5 min PBS-Ca²⁺

8. Mounting in Mowiol with 2.5% DABCO (overnight, 22°C, in dark)