Sheath fluid for Calibur summary and thanks

From: Mara Rocchi (roccm@blackie.mri.sari.ac.uk)
Date: Fri Mar 31 2000 - 10:47:07 EST


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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