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
-----------------------------------------------------------------------------
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
and distributed free of charge as an educational service to the cytometry community.
If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
