does anybody have a recipe for flow cytometry sheath fluid?

Haviland, David L via methods%40net.bio.net (by David.L.Haviland from uth.tmc.edu)
Tue Nov 30 09:39:34 EST 2010


Mary:

If this is for an analyzer, you don't, but it's a good idea for cleanliness sake.   All instruments I've worked with have an inline 0.2 or 0.45 micron filter.   Again, my comments about using DI are for analyzers ONLY.    I don't filter my saline, honestly, because I go through it so fast - I'll "burn" 20L in two days.   I make a 20X stock and go from there. 

If you and others are talking of a sorter, then yes, you must use sheath containing NaCl or you won't get any side streams and if home made is used then it should be filtered.    In that case, for my sorters I get my investigators to buy filtered sterile blood bank saline from Fischer or IsoFlow from Coulter.    I flush the entire system, filters included with 70%, and then flush it out again with the sterile saline. 

Hope that helps,

David

________________________________________
From: methods-bounces from oat.bio.indiana.edu [methods-bounces from oat.bio.indiana.edu] On Behalf Of Mary [giemsacyto from gmail.com]
Sent: Tuesday, November 30, 2010 04:23
To: methods from magpie.bio.indiana.edu
Subject: Re: does anybody have a recipe for flow cytometry sheath fluid?

On Nov 26, 9:36 am, "Haviland, David L" <David.L.Havil... from uth.tmc.edu>
wrote:
> As I run a flow core, I also use just plain old 0.9% saline as well, but... I know of other flow cytometry centers that use DI as sheath.   I B.S. you not... For the short duration of laminar flow, the central stream of sample fluid (whatever your cells are in) is centered within the DI sheath.    No one really cares what happens to the cells once past the interrogation point, let alone the waste line.   DI also tends to keep the machines cleaner and a lot less crystallization around the SIP tube.
>
> Pointed question:  Have I made the switch?   Not yet.   I have about 60L of 10X sheath to use up, but a colleague across the way uses DI, and I'm going to switch after that 60L is gone.
>
> David Haviland, PhD
> UTHSC- Houston Stem Cell Flow Center.
>
> ________________________________________
> From: methods-boun... from oat.bio.indiana.edu [methods-boun... from oat.bio.indiana.edu] On Behalf Of DK [d... from no.email.thankstospam.net]
> Sent: Wednesday, November 24, 2010 19:11
> To: meth... from magpie.bio.indiana.edu
> Subject: Re: does anybody have a recipe for flow cytometry sheath fluid?
>
> In article <de1ec71b-61bc-496f-979e-43a8b78b9... from u9g2000pra.googlegroups.com>, Mary <giemsac... from gmail.com> wrote:
>
> >Hi All,
> >> the liquid stream (sheath fluid), which carries and aligns the cells
> >> so that they pass single file through the light beam for sensing in
> >> flow cytometers does anybody knowhow to make a DIY sheath fluid? I
> >> thought maybe using 0.9% saline solution would work...amy thoughts?
> >> regards
>
> Pretty much. A simple PBS is what most everyone uses. E.g., here
> is 1X composition:http://www.biosure.com/rpages/solutions.htm
>
> Make it 10X and filter through 0.2 micron and it won't go bad even
> when non-sterile and without preservatives.
>
> DK
> _______________________________________________
> Methods mailing list
> Meth... from net.bio.nethttp://www.bio.net/biomail/listinfo/methods

David why do I need to filter sterilise the PBS?
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