I do human Monocytes all day here, In most of my studies a 100ul wb blood sample can easily yield 1000 mono's in about 7-10 min from a lysed sample using a FSC trigger. for a whole blood sample ( no lyse) using a flour trigger instead of the FSC and dilute 1/20 (to reduce coincidence) and you can get 1000 in about 10 min on LOW flow rate.... A 100 event file will contain about 2% noise ( more if whole blood)..this drops after 300 events...statistically around 1000 is a safe bet. -----Original Message----- From: bunny [mailto:bunny@cotleur.com] Sent: Monday, April 10, 2000 5:51 PM To: cyto-inbox Subject: analysis of few cells 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 9500 Euclid Avenue *+* 330-483-4800 Cleveland, OH 44195 +*+ bunny@cotleur.com 216-444-1164 *+* cotleua@ccf.org +*+ ******************************************************* When you do something, you should burn yourself completely, like a good bonfire, leaving no trace of yourself. (Shunryu Suzuki)
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