Looking for advice on pediatric samples
bunny at cotleur.com
Tue Jun 21 16:14:13 EST 2005
I'm looking for advice from someone/anyone who is currently analyzing
small cell numbers by flow cytometry. (NOT low frequency, but very few
events to begin with).
We are planning to start analyzing pediatric CSF samples in the near
future. Since we are expecting very SMALL sample volumes with very FEW
cells, I'm looking for advice on the best way to simultaneously
quantify chemokine receptors, cytokine expression and cell phenotypes.
Is flow cytometry the best method in this case? With very few TOTAL
cells, should we try a microscopy approach? Can someone point (or
SHOVE) me towards the best method?
Our current equipment choices:
At the moment, our Flow core is only equipped with a Facscan and a
LSR1. I could feasibly do 4 color (with Viability on a 5th detector,
We are waiting to find out if we will be funded for an Aria. In any
case, we could actually take samples across town to be run on an Aria.
(I'm thinking in terms of maximizing # colors at this point).
We will also have a small amt of equipment money- enough for a
FacsArray-type cytometer. Is there an advantage (as far as running
smaller samples) vs the traditional continuous stream? (We've removed
the droplet containment on the LSR1 years ago, so we already fixed the
problem with the DCM sucking the sample before we get the bar pulled
over. I regularly run 100ul samples without a problem).
Our institution also has a well equipped Confocal /Fluorescent
Thanks for any input!
Bunny Cotleur, M.S.
Sr Research Technologist, Dept of Neurosciences
Scientific Consultant, Flow Cytometry Core
Cleveland Clinic Foundation
Lerner Research Institute, NC30
9500 Euclid Avenue
Cleveland, OH 44195
Caminante, no hay camino. Se hace camino al andar.
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