I agree with Akos on both points. The Cytek AMS has been a great tool for us since ours arrived last spring and has made it possible for us to do large-scale experiments routinely. And it would be a big help to everyone if its software could be integrated with software running the cytometer. Laird Bloom Phylos, Inc. 128 Spring St. Lexington, MA 02421 > ---------- > From: Akos_Szilvasi@biogen.com > Sent: Thursday, November 8, 2001 9:53 AM > To: Cytometry Mailing List > Subject: Re: FACS Calibur + automation > > > I'd like to reply to Ray's e-mail with a quick note on automation. > > Now we have been using the Cytek 96 well plate autosampler (called AMS) > for a few months with excellent results. The instrument is reliable, fast > and no steep learning curve or training is necessary to operate it. (The > AMS is running plates right now so I can write this message instead of > hunching over the Calibur.) The wash cycle between samples can be reduced > to a few seconds because the short sample line does not retain large > volume of cells. As a matter of fact a few seconds acquisition delay > prevents contamination better than long wash periods. Practically you can > go from the end of one sample to the start of the next one in 10-12 > seconds with no contamination. The hardware is almost maintenance free and > you don't need to be present or supervise the AMS. > > I only can hope that one day manufacturers start cooperating with each > other to the benefit of their customers and saving large development > expenses for themselves. Great hardware design combined with software > integration would be a winner. I am appealing to all cytometer > manufacturers here. Take a look at the AMS. > > Akos (Biogen) > > > > > > > "Ray Lannigan" <lannigan@tritechinc.com> > > 11/07/2001 09:00 AM > Please respond to "Ray Lannigan" > > To: Cytometry Mailing List > <cytometry@flowcyt.cyto.purdue.edu> > cc: > Subject: Re: FACS Calibur > > > > > Hi Carol, > There is an alternative to the 96 well plate autosampler that BD > offers. > It is called the Automatic Microsampler system and is made by Cytek > Development. If you would like more information please contact me. > As far as sorting on the Facscaliber, recovery of and viability of > sorted cells can be an issue. The inherent problem with the mechanical > sorting technique used in the FacsCaliber, is fluid constantly flows to > your > sort vessels, even when it is not trying to sort a cell in the sort gate. > After two hours of sorting, your 1000 - 2000 targeted cells could be in > approx. 300mL of fluid. The sorting mechanism is a catcher tube that goes > into the core stream of cells and attempts to catch the cell of interest. > When a cell is moving at a velocity of 6 meters/sec forcing a small tube > into its path, then forcing it to go through that tube can put relatively > extreme shear forces upon it. Stream-in-air with the electric charge > sorting > is the way to go. > Ray > > > -----Original Message----- > From: Carol Mazurek <mazurek@mpi.com> > To: cyto-inbox Date: Tuesday, November 06, 2001 6:44 PM > Subject: FACS Calibur > > > > > >I realize that the FACS Calibur multiwell autosampler is fairly new, but > >is anyone actively using one? Could you please share your successes and > >failures with it? Does it do what you need? What would you change > >about it? Would you recommend it to others? > > > >Is there anyone with experience at sorting cells using the FACS > >Calibur? I realize it has limitations, and I'm not expecting FACS > >Vantage-quality sorting. I have a cell line that is expressing a > >recombinant protein and I want to sort the highest protein expressors > >(e.g., top 1% or 10%) in that population. What is the likelihood that I > >could get 1000 - 2000 targeted cells at >98% purity after sorting for an > >hour or two? > > > > > >Carol Mazurek > >Millennium Pharmaceuticals, Inc. > >mazurek@mpi.com > > > > > >
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