sheath fluid for Calibur

Joe Trotter jtrotte1 at san.rr.com
Wed Mar 29 23:06:44 EST 2000


Don't forget that the refractive indices of the sheath buffer and sample buffer
need to be close for good optics. Otherwise, you have what amounts to a dynamic
lens at the water/saline interaface interfering with the light collection. With
strong signals as in most assays for DNA in mammalian cells, the loss of CV is
probably not too serious. With yeast, for example, matching buffers is more
essential.

   My son did a science fair project on this very issue several years ago. He used
glutaraldehyde fixed Chicken Red Blood Cells suspended in either water or in
saline. He ran both on a FACS Calibur with 1) water as a sheath buffer, and 2)
saline as a sheath buffer. He then compared CVs and the quality of the data. Try it
yourself and draw your own conclusions.

            Joe

Zucker.Robert at EPAMAIL.EPA.GOV wrote:

> The question on sheath fluid is whether inexpensive water is equivalent to
> expensive saline. We use filtered water on a FASCalibur primarily for DNA
> analysis.  We have not seen any problems due to possible mixing of sample with
> sheath. Using Salt has one potential problem. Salt must be cleaned from the
> system or it can cause some major  fluidic problems, namely blockage (minor and
> major). Water is easier and cheaper to use.
> Bob
>
> Robert M. Zucker, Ph.D
> U.S. Environmental Protection Agency
> National Health and Environmental Effects Research Laboratory
> 2525 NC Highway 54,  MD 72
> Research Triangle Park, North Carolina, 27709
> tel: 919-541-1585 fax 919-541-4017
> e-mail: zucker.robert at epa.gov
>
> |--------+----------------------->
> |        |          lannigan at trit|
> |        |          echinc.com   |
> |        |                       |
> |        |          03/28/00     |
> |        |          08:40 AM     |
> |        |          Please       |
> |        |          respond to   |
> |        |          lannigan     |
> |        |                       |
> |--------+----------------------->
>   >-------------------------------------------------------|
>   |                                                       |
>   |       To:     cytometry at flowcyt.cyto.purdue.edu       |
>   |       cc:                                             |
>   |       Subject:     Re: sheath fluid for Calibur       |
>   >-------------------------------------------------------|
>
> Mara,
>     Since the fluid systems on the Facscan and FacsCaliber are functionally
> similar, using the same sheath fluid for both cytometers should be OK.
> Ray Lannigan
> Tritech Inc.
> 800-886-7004
> www.tritechinc.com
> -----Original Message-----
> From: Mara Rocchi <roccm at blackie.mri.sari.ac.uk>
> To: cyto-inbox
> Date: Monday, March 27, 2000 4:16 PM
> Subject: sheath fluid for Calibur
>
> >
> >Dear Flowers,
> >we are in the process to buy a FACSCalibur, and we would like to here your
> >suggestion abut the kind of sheath fluid we must/can use. We aren't
> planning
> >any sorting, maybe some DNA work, lot and lot of phenotyping and
> >intracellular staining.
> >>From the archives looks like that different people use different
> solutions,
> >and for different reasons. I have been using sterile saline for FACScan and
> >FACStar, and D'PBS for FACSort, but being new to the Calibur world, I would
> >like to be sure that those alternatives to the BD sheath fluid are OK.
> >
> >Thanks a million in advance
> >Mara
> >
> >Mara S.L. Rocchi
> >Moredun Research Institute
> >Penicuik EH26 0PZ
> >Scotland, UK
> >e-mail: roccm at mf.mri.sari.ac.uk
> >




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