does anybody have a recipe for flow cytometry sheath fluid?
Mary
> 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
Haviland, David L
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...@oat.bio.indiana.edu [methods...@oat.bio.indiana.edu] On Behalf Of DK [d...@no.email.thankstospam.net]
Sent: Wednesday, November 24, 2010 19:11
To: met...@magpie.bio.indiana.edu
Subject: Re: does anybody have a recipe for flow cytometry sheath fluid?
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
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Mary
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.
>
> ________________________________________
> Sent: Wednesday, November 24, 2010 19:11
> Subject: Re: does anybody have a recipe for flow cytometry sheath fluid?
>
>
> >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
David why do I need to filter sterilise the PBS?
Haviland, David L
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...@oat.bio.indiana.edu [methods...@oat.bio.indiana.edu] On Behalf Of Mary [giems...@gmail.com]
Sent: Tuesday, November 30, 2010 04:23
To: met...@magpie.bio.indiana.edu