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We need to do intracellular calcium levels in yeast cells and it would save
a lot of time & trouble if someone could provide some tips about indo-1
loading. For instance, do you need to use protoplasts and are there any
cell membrane pumps that need to be inhibited for proper loading? We
haven't tried indo yet but I couldn't detect any fluo-3 loading on my first
attempt.
We would also like to do cell cycle measurements and would be very grateful
if anybody could provide a protocol for uniform permeabilisation of the
cells. My first attempts using freeze-thaw, at the suggestion of our
molecular biologists, were unsuccessful because of incomplete dye
penetration.
We would welcome any suggestions,
Dean R. Hewish, Cell Biologist & Flow Cytometrist.
CSIRO Biomolecular Engineering, 343 Royal Parade, Parkville, 3052,
Victoria Australia.
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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;
Web