Thanks a million to everybody that answered my query about what to use as sheath fluid for the Calibur. Having had many direct replay I decided to post a summary of the answers. Here it is: First of all not many people use proprietary sheath fluid. Wonder why? Here the favourite ones: Water: many many flowers happily use filtered water, is cheap and keep the machine very clean and the system noise-free; just remember that with water the FACS probe in the tank will thinks that the tank is empty, and you will have to cheat and disable the probe itself (with the help of a little conductive foil). Looks like that the difference in the refractive index between water and your sample fluid could increase your noise (Alice). Saline: cheap and easy, but can cause problems of blockage if you don't clean the salt from the system (regular cleaning procedure in place). Home-made PBS: many flowers use it as well, sterile for sorting, and always well filtered to reduce the background noise. If you have a central core service or a pharmacy they can make it 10x for you, and you can dilute it with milli-Q water when needed, saving space as well as money. Commercial buffers: they are expensive, but often cleaner than home-made PBS (apparently when you go from PBS to a commercial buffers your nice tail of low FCS debris will disappear). You can buy bulk quantity of fluid from companies that don't sell cytometers and save money. Other suggestions were that 1) the refractive indices of your sheath buffer and sample buffer should be as close as possible for good optics; 2) if you use water or a home-made saline or PBS rinse your machine with commercial buffer that contains preservatives before the week-end if the machine is not used; 3) for sorting looks like that almost everybody goes back to commercial fluid (without preservatives!) or sterile PBS. Thanks a lot again, we made our decision! Cheers, Mara Mara S.L. Rocchi Moredun Research Institute International Research Centre Pentland Science Park Bush Loan Penicuik EH26 0PZ Scotland UK e-mail: roccm@mf.mri.sari.ac.uk
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