FACSCalibur limit of detection analysing bacteria

From: Daniel Hoefel (daniel.hoefel@sawater.sa.gov.au)
Date: Sun Mar 04 2001 - 20:41:15 EST


I am interested in enumeration and identification of bacteria from water samples
using flow cytometry.  I recently attempted to determine the limit of detection on
a FACSCalibur for enumeration of E.coli cells suspended in MilliQ water (filtered
distilled water). The experiment was carried out on an overnight broth culture culture
of E.coli stained with SYTO9.  MilliQ water was used as sheath fluid.  I was able to
get down to determining  the concentration of 500 E.coli cells/ml where below this
number results became inaccurate.  I was looking to enumerate down to about 100 E.coli
cells/ml and I was wondering if anyone has done this type of experiment before and if
they were able to accurately detect less than 500 cells/ml. Am I asking too much from
the FACSCalibur and/or are there any secrets to improving the limit of detection?

Input from anyone on this matter would be greatly appreciated.

Daniel Hoefel
Email: daniel.hoefel@sawater.sa.gov.au



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