bacteria by flow
mlunzer at cbs.umn.edu
Tue Nov 15 14:02:27 EST 2005
I run E. coli in a FACScalibur all the time. The trick is to have the
machine clean and run on the lowest setting. To clean the machine I run
straight bleach for 5 min followed by 5 tubes of 0.2 um filtered PBS.
This is all done in the carousel. When you get to the last tube you
should see very few counts (15 events per second every now and then).
I use YO-PRO-1-iodide to stain cells that have been treated with T5
bacteriophage, chloramphenicol, and EDTA. You could probably use my
settings as a guide, so here they are,
FSC E01 Log
SSC 415 Log
FL1 505 Log
FL2 477 Log
FL3 568 Log
Trigger on both FSC & SSC
For the first time we are thinking to run bacteria in our FACScalibur.
Is there someone that can give me some good advices? Basic questions
like: 1. it is ok to resuspend in normal FACS buffer (FCS, azide) 2.
which would be the best instrument settings to get started (mostly FSC
and SSC). We want
to look at cultured e-coli/eGFP and fecal samples which should contain
the same bugs. 3.
cell concentrations? 4. Special cleaning advices?
Thanks in advance
Corrado M. Cilio, M.D., Ph.D. Assistant Professor Cellular Autoimmunity
Unit Dept. of Clinical Sciences/Paediatrics Malmo University Hospital
Lund University 205 02 Malmo, Sweden
More information about the Cytometry