http://confocal-microscopy-list.275.s1.nabble.com/Suggestions-for-4-colour-stain-on-Leica-SP5-tp3396389p3460942.html
Thanks for the suggestion Arne. Have you used this Ab for marking plasma membranes for segmentation before? What has been your experience with this reagent?
Samuel A. Connell
St. Jude Children's Research Hospital
against the alpha subunit of the NKAtpase. It's called alpha5 (a5).
> Tim,
>
> Do you have a particular Ab to recommend for the sodium-potassium ATPase you mentioned?
>
> Thanks,
>
> --
> Samuel A. Connell
> Director of Light Microscopy
> Cell & Tissue Imaging Center
> St. Jude Children's Research Hospital
> 262 Danny Thomas Place
> Memphis, TN 38105-3678
> Office (901) 595-2536
> Cell (901) 603-3162
>
[hidden email]
> ________________________________________
> From: Confocal Microscopy List [
[hidden email]] On Behalf Of Timothy Feinstein [
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> Sent: Thursday, August 06, 2009 8:41 PM
> To:
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> Subject: Re: Whole Cell Dye. .
>
> Hi Cam,
>
> In the past we have had good results with chloromethylfluorescein
> diacetate (CMFDA). CMFDA is an intravital dye that fluoresces in the same
> channel as GFP, however as far as I know only works in live cells. If
> transfection is possible, people often use GFP-family proteins as a
> co-transfection marker. These pass freely through nuclear pores and
> therefore produce even cellular labeling. If an antibody is preferable,
> the sodium-potassium ATPase marks the plasma membrane as clearly and
> evenly as anything that I have seen.
>
> All the best,
>
>
> Tim
>
> Timothy Feinstein, PhD
> Postdoctoral Associate, Vilardaga laboratory
> University of Pittsburgh Dept. of Pharmacology and Chemical Biology
> Pittsburgh, PA
>
>
>
>> 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
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>> Royal Melbourne Hospital
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>>
>>
>>
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