Re: Cell Sizes, FSC, and SSC

From: mail.thelinks.com (sstouden@thelinks.com)
Date: Tue Jul 02 2002 - 06:55:45 EST


The question still begs for information on the relative cell sizes by cell
type, and organelle content counts.  What means of gathering this data
exists and who compiles it?  Has it been done for most cell lines?

Actually, the data needed is the ontological development of the cell (by
type, organelle counts and external signal effectors)from its stem cell
origin to its quantized fully developed mature cell type both in vivo and
in vitro.

In my opinion, this data is so important that it should be a national
initiative by one or more of the government agencies and it should not be
limited to animal cells but should also include plant cells.

sterling

At 08:12 PM 6/30/2002 -0400, Howard Shapiro wrote:
>
>Annette Byrne wrote:
>
> > Hello all
> >
> > Does anyone have an info on relative sizes of the following cell types:
> > Fibroblasts, endothelial cells, smooth muscle cells and macrophages. I am
> > trying to make a gross distintion of a mixed cell population containing
the
> > cells above based on their FSC/SSC.
> >
> > If anyone could order them based on largest to smallest cell type I would
> > be eternally grateful !
> >
>
>FSC does not measure cell size!!!!!!!! In some instruments, 5.5 um plastic
>beads have bigger FSC signals than 5.0 um plastic beads (and in other
>instruments, they don't).  These beads are relatively uniform and
>smooth-surfaced and spherical, and all have pretty much the same refractive
>index; FSC (and SSC) signals are influenced by both size and refractive
>index. The refractive indices of cells, by contrast, vary with the cells'
>content of water and of proteins and other macromolecules. If you can't
>reliably use FSC to tell that a large lymphocyte is bigger than a small
>one, you certainly can't use FSC to compare relative sizes of different
>cell types. If you happened to have access to pure populations of the cell
>types you were interested in, you could produce FSC vs. SSC plots for each,
>which might help in resolving a mixture, provided the cells in the mixture
>had not been exposed to conditions that could change their refractive
>indices or sizes.
>
>The best way to identify mixed cell types on FSC vs. SSC plots is to gate
>individual cell types based on the presence of a specific marker, but you
>have to be sure of the marker specificity - see my reply to a posting from
>Ray Hester on reticulocytes.
>
>And, as far as sizing goes, while Coulter volume has its problems, it's far
>more accurate than scatter measurements.
>
>-Howard
>
>
Freedom is "unbounded, ungated option" to engage tangible and intangible
human experience within the space defined by the farthest extent of
individual imagination.   s. stoudenmire, 6.22.2002



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