Hi Rochelle
Yeast tends to flocculate. As the step is Ca dependent EDTA
helps. Certain sugars can also prevent their lectin
interaction. My other recommendation is to actually install
a 50um nylon filter straight on the sample insertion rod as
aggregations are very unlikely to occur once they are in
there.
I can't comment on the disinfection as I chucked the bit of
sample tubing after running 3ml of 50% Domestos (UK
formulation with bleach).
The GFP signal we looked at was too strong to note
autofluorescence interference.
Good luck
Gerhard
______________________________ Reply Separator _________________________________
Subject: sorting live yeast
Author: diamond@cco.caltech.edu at INTERNET
Date: 07/01/98 17:35
Hi Flow Community-
I would like to get some information about sorting live yeast cells.
In particular I would like to know the following:
How badly do they reaggregate after filtering thru nylon mesh ( do
I need to worry about the size of my orifice?)
How do you decontaminate the sorter after the run (preferred
cleansing agents)?
What kind of autofluorescence do yeast cells (S. Cere.) exhibit?
Any experiences good or bad would be useful.
The experiment is to sort out high and low expressing GFP (EGFP-like)
cells.
Thanks ever so much
Rochelle Diamond
Caltech Biology
Flow Cytometry/Cell Sorting Facility
diamond@cco.caltech.edu
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