Sorry, let me correct that last post Tim, if you triggered off alexa 568 then you would only be looking at the positive, and you need to discriminate from negative. So I guess you need to trigger off side scatter and then look at the fluorescence. Good luck and let me know if it works! Cheers, Eric Eric O'Connor Head of Flow Cytometry MRC Clinical Sciences Centre Imperial College Faculty of Medicine Hammersmith Hospital Campus DuCane Road, London W12 0NN Tel: +44(0) 208 383 8330 Mobile: +44(0) 781 575 7730 ________________________________ From: O'Connor, Eric Sent: 27 June 2008 12:48 To: cyto-inbox Subject: RE: Particle sorting request I'm not certain the size of the bacteria we used to sort but I don't think they were much bigger than 200nm and we could pick them up fine triggering off side scatter on a Vantage SE. Your other option depending on your instrument could be to trigger off of Alexa 568 and then look at side scatter. If this is something you want to do long term and have a decent sized budget, you might want to talk to Cytopeia in Seattle who custom make cytometers for analysis of cyanobacteria and other tiny sea critters involved in carbon cycling. Cheers, Eric Eric O'Connor Head of Flow Cytometry MRC Clinical Sciences Centre Imperial College Faculty of Medicine Hammersmith Hospital Campus DuCane Road, London W12 0NN Tel: +44(0) 208 383 8330 Mobile: +44(0) 781 575 7730 ________________________________ From: Timothy Overton [mailto:two812@bham.ac.uk] Sent: 26 June 2008 15:17 To: cyto-inbox Subject: Particle sorting request Dear All, I just has a request from someone working in my department which I think is theoretically possible, but I wondered if anyone has tried anything like it. They have nanoparticles coated in biotin, to which are attached Streptavidin conjugated to Alexa 568. The size of the particles is around 200 nm across. They asked whether flow cytometry could be used to determine the proportion of particles with Strep-Alexa attached, and then if they could be sorted. Has anyone done anything like this? Thanks, Tim ***************************************** Dr Tim Overton Lecturer in Biochemical Engineering School of Chemical Engineering The University of Birmingham Birmingham B15 2TT t: +44 (0) 121 414 5306 e: t.w.overton@bham.ac.ukReceived on Fri Jun 27 17:49:09 2008
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