Re: Whole Cell Dye

Posted by Cameron Nowell on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Suggestions-for-4-colour-stain-on-Leica-SP5-tp3396389p3428329.html

Thanks Michael. Nice paper but unfortunately it only deals with single
cells that are nice and flat. We are trying to segment confluent layers
of cells that are fairly tall.




-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of MODEL, MICHAEL
Sent: Wednesday, 12 August 2009 12:21 AM
To: [hidden email]
Subject: Re: Whole Cell Dye

A paper has been published on this subject: "Comparison of reagents for
shape analysis of fixed cells by automated fluorescence microscopy",
Cytometry 52A, 90-100, 2003



-----Original Message-----
From: Cameron Nowell [mailto:[hidden email]]
Sent: August 6, 2009 7:38 PM
To: [hidden email]
Subject: Whole Cell Dye

Hi List

Can anyone recommend a dye (or antibody) that will stain the whole cell
evenly to the plasma membrane? I need a dye that will give fairly even
staining across the whole cell to make segmentation easier.

I have tried CellMask but is it a bit variable in its staining pattern,
there are blobs in the cytoplasm and nucleus may or may not stain up.
Something that stains just the plasma membrane would also be suitible. I
have used the invitrogen FX dyes before (not for this experiemnt) and
have found them a bit hit and miss too. I have also tried WGA but it
doesn't give an even stain around the mebrane which makes segmentation
quite dificult.

Any suggestions would be greatly appreciated.


Thanks

Cam


Cameron J. Nowell
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008
Royal Melbourne Hospital
Victoria, 3050
AUSTRALIA
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