Re: Whole Cell Dye

Posted by axel k preuss on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Suggestions-for-4-colour-stain-on-Leica-SP5-tp3396389p3428501.html

How about the live cell stains like SYTO (Mol Probes). Membrane stain would be R 18?
-axel

--- On Tue, 8/11/09, Cameron Nowell <[hidden email]> wrote:

> From: Cameron Nowell <[hidden email]>
> Subject: Re: Whole Cell Dye
> To: [hidden email]
> Date: Tuesday, August 11, 2009, 11:33 PM
> 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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>
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