Antwort: consistency in counting cells by flow

Rick Arthur Bright rbright at emory.edu
Wed Mar 20 20:16:17 EST 2002


BD does sell the Trucount tubes alone without the kit.  Some of the sales
reps and customer service folks say no at first, but if you tell them to
check  again or ask someone, they will find out that they are available
alone.  I buy them every month to use with mouse reagents.


On Tue, 19 Mar 2002 eug.kreysch at t-online.de wrote:

>
> Hi Yoav,
> we have used Becton Dickinsion´s TruCount beads (defined number of beads in
> FACS tubes, see http://www.bdbiosciences.com/immunocytometry_systems/) with
> good results and calculated absolute cell numbers from total events (e.g.
> 10,000 to 15,000) using cell and bead counts within the mixture after
> gating on beads and PI (propidium iodide) negative live and/or PI positive
> dead cells mainly according to BD´s protocol. However, for an accurate
> calculation proper mixtures/proportions of beads and cells (not to many
> cells!) are necessary.
> The major disadvantage is that - as far as I know - BD does not offer the
> beads separately under normal conditions. Nevertheless I would ask .....
> Best regards
> Georg Kreysch
>
>
>
>
>
> yaltman at liai.org am 15.03.2002 23:52:01
>
> An:   cytometry at flowcyt.cyto.purdue.edu
> Kopie:
>
> Thema:    consistency in counting cells by flow
>
>
> Dear Flowers,
>
> A colleague of mine wishes to use a FACScan to count cells for an
> experiment.  The experiment will proceed over several weeks, and he
> wishes to count live and dead cells over several time points (eg. 1
> wk, 2 wks, etc.)
>
> Assuming that his samples are resuspended in the same volume every
> time, and that the flow rate would be the same from week to week or
> sample to sample, he could just take the number of events in his live
> gate vs. non-debris events.  But, he is concerned that flow rate may
> vary from week to week due to small obstructions in the fluidics.
>
> It seems that if we had a bead of known concentration we could add
> these to the sample and do something like stop collecting after
> 10,000 beads and thus know we had the same volume as last time.  Does
> this seem like a viable approach.  Does anyone make beads in known
> concentration, or is there some other standard we can use to know
> that we are counting cells in the same volume from week to week.
>
> Thanks,
>
> Yoav
> ---
> Yoav Altman
> Imaging Facility Specialist
> La Jolla Institute for Allergy & Immunology
> 10355 Science Center Drive
> San Diego, CA 92121
>
> email: <yaltman at liai.org>
> Phone: (858) 678-4608
> Fax: (858) 558-3526
>
>
>
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>




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