> I would like to get the group's opinion on the best available >instrument for doing multicolor [ at least 6, in the near future maybe >7, and potentially 8 ] analysis as to >Altra vs Vantage vs Moflo > >1] how well does the 8x8 compesation matrix on the Moflow work? > >2] if I need to use aftermarket software compensation , is there any >advantage to Mac based data vs. Windows based data? > >3] is there a big difference in sensitivity between the jet in air flow >cell [Moflo &BD] and the quartz [BD & Coulter] for reagents such as >Cascade Blue and APC-CY7? > >4] which has the best design for multilaser alignment? > >5] which has the best disign and optics for signal collection? > >Thanks for any info. > >Regards, >Tom > >-- >***************************************************************************** >Thomas W. Mc Closkey, Ph. D. >Director of Flow Cytometry, North Shore University Hospital >Assistant Professor of Pediatrics, New York University School of >Medicine >Boas Marks Biomedical Research Center, 350 Community Drive >Manhasset, Long Island, New York 11030 >ph: 516-562-4844 [office], 516-562-1135/4641 [lab] fax: 516-562-2866 >***************************************************************************** Hi Tom, I'll try to answer your questions one at a time where I can. 1] how well does the 8x8 compensation matrix on the MoFlo work? -- The compensation matrix is quite good on the MoFlo, there is a little distortion of the populations due to the algorithm, but it is really of no significance. I built analogue compensation circuits for my MoFlo couple of years ago because; a) The software compensation algorithm on the MoFlo was dreadful. It has much improved and they have moved the compensation matrix onto a DSP chip that now allows you to sort on compensated parameters. b) Then, you couldn't sort on compensated parameters. Although my analogue electronics worked well it did have a few problems with phase alignment of pulses which was exacerbated at high signal input into the compensation. And it was not capable of interbeam compensation. Cytomation, in utilizing a DSP chip and improving their algorithm, have sorted their problems out. The DSP has a number of advantages over the analogue solution. The DSP does complete interbeam compensation. DSP outputs are saved along with the uncompensated data allowing you to go back after acquiring your data and re-adjust the compensation. The overhead is that your file size is doubled, but that is no real pain when you have 12GB at your disposal. The DSP has no pulse phase problem. 2] if I need to use aftermarket software compensation , is there any advantage to Mac based data vs. Windows based data? -- I'm keeping out of this one... I like Acorn RISC OS, but that's another thread! 3] is there a big difference in sensitivity between the jet in air flow cell [MoFlo &BD] and the quartz [BD & Coulter] for reagents such as ascade Blue and APC-CY7? --I haven't used these reagents so I cannot comment. However, I ran some really ancient 5-peak calibration beads to compare sensitivities of my MoFlo and my FACSCalibur (running at 35 psi with 100u nossle) I've enclosed the results. The MoFlo was better but I do think that my BD machine needs realigned. 4] which has the best design for multilaser alignment? -- In my mind MoFlo for high speed sorting, because each beam is independently steered and focused. I can't comment on the Altra as I have never seen one running, but I think they use independent beam steering and focusing (please correct me if I'm wrong). 5] which has the best design and optics for signal collection? -- Again I would say the MoFlo, because the optical bench, electronic rack and the collection optics are open so you can easily add more PMTs, ADCs, etc,. I hope this helps. Best Regards, Andy.
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