RE: Re: Keeping large cells in suspension (Methyl Cellulose)
john michie (jm5@maties.sun.ac.za)
Tue, 15 Aug 95 12:52 +0200
> Parasitologists have long used methyl cellulose to slow down single cell
> organisms such as ameobae, flagellates etc. for microscopic study. In
> fact such a product was (is?) commercially available (I don't know from
> whom) under the name "Proto-Slo". The significance of this is that these
> eukaryotic organisms stayed alive during the time they were suspended in
> this material. I seem to remember (but I also forget alot) that this
> solution was about 8% methyl cellulose. Hope there is grain of
> usefulness here for suspending your particles. The other thing that
> comes to mind is 0.5% agarose if you can maintain a temperature above the
> geling point.
> ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
>
> Steve Kayes
>
> On Wed, 9 Aug 1995 gap@mit.edu wrote:
>
> > Hi All!
> >
> > Has anyone found a way to keep large particles in suspension for
> > any extended period of time? The problem I am having is that I am
> > trying to sort a fraction of 100um particles and these things settle out
> > of solution after only 10 minutes or so. This makes the sort long and
> > tedious. I once heard about a food additive called xanthan gum that
> > would increase the viscosity of the liquid the particles are in. Does
> > anyone have this or another product working?
> >
> > Thanks in advance.
> >
> > Glenn
> > MIT Flow Cytometry Core Facility
> > (617) 253 6454
> > gap@mit.edu
> >
> ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
> I have used methyl cellulose for plating out primary human tumour cells into
> multiwell plates, where there was the need to slow down the settling rate so
> as to obtain an even distribution of cells on the floor of the wells.
> Concentration used was 6%. It worked well. Methyl cellulose is more soluble
> at low temperatures, so there is a somewhat lenghty protocol for preparation
> ofsterile solutions (if anyone wants it, let me know).
> Whether methyl cellulose is something you want to run through your flow
> cytometer is another thing though. If you look at a solution of methyl
> cellulose under an inverted microscope, you will see that it is a suspension
> of small pieces of cellulose. I would think that this would create a lot of
> "noise" in a FSC vs SSC plot.
John Michie PhD "Life is too short to
Department of Radiation Oncology drink bad wine"
Gene Louw Building
Tygerberg Hospital
Cape Town 7505
South Africa
E-Mail: jm5@maties.sun.ac.za
Tel: 027 21 938 4911 bleep 348
Fax: 027 21 931 0804
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