Hello All!
        I have a question regarding fixation of tissue cultured cells. When fixing
with 4% paraformaldehyde (10 min, room temp), the cells appear to shrink. I
realize this is a characteristic of this kind of fixation, but it really
messes up the morphology of cell-cell junctions. Methanol or acetone, or
the combination (-20, dry) doesn't seem to work either. I've tried
formaldehyde as well. I've also tried a cytoskeletal stabilizing buffer
(50% glycerol) that improved the morphology, but the cells still appear
stringy and pulled apart, instead of nicely abutting each other. We plate
our cells on a collagen/fibronectin matrix, which dramatically improved the
morphology, but it still lacks the pre-fixation appearance of confluent
cells. We have followed published protocols in papers about cell
junctions, but it just isn't working for us. Any suggestions?
        Thanks!
        Kathy




Kathy Spencer
The Scripps Research Institute
10550 N. Torrey Pines Road
IMM 24
La Jolla, CA 92037
(858) 784-9372
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