> 3. Can the Hoechst emission spectrum be manipulated to shorter
>wavelength in any way (we have tried a longer FITC filter, 535DF15, but
>that didn't help much)? BTW, our Hoechst fluorescent beads from Flow
>Cytometry Standards give *much less* spillover!?
>
Do the beads have Hoechst bound to DNA or just a high concentration of Hoechst?
>
> 4. Are either of 33342 and 33258 superior in this context?
>
Probably not with fixed cells.
> 5. Is this experiment *obviously* impossible?
>
No, but it is at least difficult. Why do you need Hoechst in particular if
the cells are fixed, or are you constrained from using RNAse? And why is
not separating the beams so important?
--Howard
CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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