Hi Nathalie, The main issue is what makes a cell fragile; e.g I sort heat-shocked mosquito cells; they're in pretty bad shape by the time I get them, have a very long turnover time and don't grow very well if too dilute in culture. On account of these factors I have to run a very large volume of sample to obtain enough target cells for subsequent culturing. The high speed sorter -the MoFlo in my case is ideal due to, speed of course, obtaining a higher number of target cells and the cells get back to the incubator sooner etc. So basically if a cell is fragile, it is not neccesarily the sorter that kills it; one has to consider various factors and strategies. For any sorter; sample preparation/collection is a vital factor; e.g the presence or absence of serum in the sample, the medium the cells are collected in, whether the cells are resuspended in fresh medium after sorting, temperature etc. When high speed cell sorting; if you concentrate your cells to say 5 times the concentration you normally use on a non-high speed sorter then you also have to increase the concentration of serum. You can only concentrate adherent cells to a certain extent anyway and trying to run them in say 1%BSA/PBS will give you loads of clumps, clogs, dying cells etc. My adherent cells now arrive in medium with 10%FCS. O.K the background fluorescence is a bit higher but that doesn't affect what I want to sort. So there was a bit of trial and error here on my part. Of course with high speed sorting pressure is a factor that has to be considered, but the ideal pressure for a given cell can be found. I don't think that one can say that a certain type of sorter gives better viability than another type. Someone should correct me if I'm wrong. To answer your question I don't have any viability problems with the MoFlo -touch wood. Regards Ann At 16:40 31.07.00 -0400, you wrote > >Greetings to all flowers > > >We are going to buy a high speed cell sorter and are really considering >the Moflo. Somebody raised the issue of viability when sorting fragile >cells with the Moflo. I would like to know if any Moflo users >encountered that problem. Any information would be very usefull to us. > >thanks in advance > >-- >Nathalie Tessier, M.Sc. >Head of Flow cytometry Service >Institut de recherches cliniques de Montréal >110 west, Pine Avenue >Montreal, Quebec, Canada >H2W 1R7 >tel: 514-987-5608 >fax:514-987-5736 > > >
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