Hi Bunny, I'd suggest either setting up the instrument using a large sample available, stained and with controls identical to your actual sample and running your actual sample afterwards - no need to waste cells optimizing settings that way. Other alternative is to take your sample to a scanning microscope type cytometer; no single cell should get lost that way. I've got a FACSarray, but can't see a big advantage over a conventional cytometer in terms of number of cells required for analysis. It does use a syringe drive type of injection mechanism, so maybe a little less volume gets wasted - but you could fix that up for your sample simply by diluting it a bit more than usual anyway. Guy Next generation therapeutic <http://www.ablynx.com/> antibodies Guy Hermans, PhD Project Leader Ablynx NV Technologiepark 4 B-9052 Zwijnaarde Belgium guy.hermans@ablynx.com tel: fax: mobile: +32 (0)9 261 06 57 +32 (0)9 261 06 27 +32 (0)486 788 551 <https://www.plaxo.com/add_me?u=30065269879&v0=994426&k0=2009290972> Add me to your address book... <http://www.plaxo.com/signature> Want a signature like this? -----Original Message----- From: bunny [mailto:bunny@cotleur.com] Sent: Tuesday, June 21, 2005 11:14 PM To: cyto-inbox Subject: Looking for advice on pediatric samples Hello Flowers! 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, using UV). 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 Microscopy core. Thanks for any input! bunny 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 Lab: (216)444-1164 ********************************************* Caminante, no hay camino. Se hace camino al andar.
This archive was generated by hypermail 2.1.8 : Sat Jan 14 2006 - 22:03:48 EST
