 |
 |
 |
 |
We have used Hoechst 33342 to stain HeLa and CHO cells and sorted them
through the cell cycle. CHO cells don't stain very well with this dye,
because they tend to pump out the dye via the MDR pathway (as Krishan has
pointed out). However, using valinomycin or trifluoroperazine can yield much
better DNA histograms and still get excellent viability. Viability goes down
if the UV laser intensity is too high. We normally use 50 mW. This
procedure works well for a variety of cell types. I am not aware of
published results using DAPI instead of Hoechst, and 33342 is usually used
instead of 33258. Since both dyes are excited with the UV, there is no good
reason not to use Ho342. I can give more detailed procedures if you are
interested.
Mike Fox
Dept of Radiological Health Sciences
Colorado State University
Ft. Collins, CO 80523
Tel: 970-491-7618 FAX: 970-491-0623
|
|
|
|