Re: analysis of few cells

From: Frederic Preffer (preffer@helix.mgh.harvard.edu)
Date: Thu Apr 13 2000 - 22:14:44 EST


Dear Bunny

I would be very cautious about looking at a few cells (<100); however i am
entirely confident that this can be done by flow cytometry.. we have looked at
hundreds of csf or vitreous fluids as clincal cases over the last few years.
When done by professional/ experienced folks like those i am happy to say work
in our lab here, the results are entirely trustworthy.  The Compucytometer is
also likely able to do this type of work.

see
Kelliher AS, Lau S,  Olszak I,  Dombkowski  DM and FI Preffer FI (1999)
Multiparameter Flow Cytometry on Cerebrospinal and Vitreous Fluid Specimens:
Use in Identifying Hematologic Malignancy in the Central Nervous System.
Cytometry 38: p336 #41. CAC Conference, Palm Springs CA

F Preffer


At 05:51 PM 4/10/00 -0400, bunny wrote:
>
>Fellow Flowers:
>I wonder if I could gather some opinions on releveance as it relates to
>minimum numbers of events collected.
>I am assisting a lab in developing FACS analysis of PBMC's and monocytes
>in CSF. They informed me that when they run FACS, they are "lucky" to
>get 100 events.  I raised the questions of relevance, stating that those
>100 "events" could as easily contain some non-cell events as cell
>events. They insist their data is good becasue it compares somewhat (?)
>with a lab in Europe they are collaborating with. We have had many
>conversations on this.
>(I should mention- neither lab has direct FACS experience. They got all
>their protocols second hand, and just collect what they collect).
>Additional information: they are looking at chemokine expression
>(another difficult task) on these few cells- and I believe they are
>using the PBMC's to set up isotype controls. I doubt they even use comp
controls.
>
>Please throw your 2cents my way. And if any of you do this type of
>analysis (few cells) with alternative techniques- I'd love to hear about it!
>Thanks-
>--
>
>
>
>Bunny
>
>
>
>
>
>
>*******************************************************
>Bunny Cotleur                    +*+       Bunny  Cotleur
>Cleveland Clinic Foundation      *+*       2001 Lester RD
>Neurosciences  NC30              +*+       Valley City, OH  44280
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>Cleveland, OH 44195              +*+       bunny@cotleur.com
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>cotleua@ccf.org                  +*+
>
>*******************************************************
>When you do something, you should burn yourself completely, like a good
>bonfire, leaving no trace of yourself.
>(Shunryu Suzuki)
>
Frederic I. Preffer
Department of Pathology
Charlestown Navy Yard- 7140
Massachusetts General Hospital East
Charlestown, MA 02129

voice     [617] 726-7481
fax        [617] 724-3164



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