Re: Bacterial Counts on Flow

From: Mark Kukuruga (kukuru@med.umich.edu)
Date: Wed Sep 19 2001 - 09:57:03 EST


Amy,
The success of your analysis may depend on your knowledge of the kit used . . . if
you're using PI, that's fairly straightforward.  PI(-) cells are "alive," PI(+)
cells are "dead."   If you're using the SYTO 9 dye, you may see all cells light up,
but counter-staining again with PI should give you the same live/dead difference.
Me? . . . I'd start with the microscope - - if you can see it there, you can flow it.
Luck.
MAK.

P.S. . . . Bugs flow different from other cell types.  Typically smaller, they don't
always behave predictably in a fluid stream.  Cell concentration and core stream
diameter are critical.

--
Mark A. KuKuruga, Managing Director
University of Michigan Flow Core
7416 CCGC 0946
(734) 647-3216, fax (734) 936-7376
kukuru@umich.edu


>>> " Amy Nield" <anield@cpd2.usu.edu> 09/17/01 01:40PM >>>
Hi Flowers,

    I have a question that I hope I can get some help with.  We have a client that
    would like to use the Flow to get an accurate count on his bacterial populations.
    He has a kit that he got through Molecular Probes, that stains DNA based upon if
    the cell membrane is intact or not.  Basically he would like to count the live cells
    vs. the dead cells.       When we attempted the kit we could not get any separation
    of live vs. dead cells.  Any help would be greatly appreciated.  If anyone knows
    of this kit (BacLight Bacterial Viability Kits) and how it works please let me know.

Thank you for your help,

Amy Nield



This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:31 EST