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Dear Andrea

Thank you very much for your reply. This at least saves me from more 
succesless attempts.
I must say that then I was misinformed by the invitrogen service.

Since DiI won't work on living cells I'll repeat the last part of my former 
mail as a new subject:
Does anybody know of a red fluorescent non toxic stain for imaging the cell 
nucleus or nuclear membrane in living cells?
Thanks in advance

Winnok

----- Original Message ----- 
From: "Dr. Andrea J. Elberger" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Tuesday, September 13, 2005 3:47 PM
Subject: Re: DiI labeling


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> WINNOK - Your outcome using DiI was due to the fact that you used it in 
> live cells.
>
> Even in neurons, when DiI is applied to live cells it is endocytosed and 
> transported in the cytoplasm; in the case of neurons, it is transported 
> retrogradely to the cell body.
>
> It is only when you use the DiI in aldehyde-fixed tissue that you see it 
> as a lipophilic dye that passively diffuses within the lipid bilayer so 
> that the entire cell membrane is labeled.
>
> Sorry, but DiI can't give you the results that you want in these 
> conditions. You are right to seek another live counterstain.
>
> ANDREA ELBERGER
>
>
> winnok wrote:
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Dear all,
>>
>> I try to use DiI-solution (molecular probes) as a counterstain for the 
>> membranes of endothelial cells but usually it is visible as vesicles 
>> probably due to endocytosis.
>> I have tried several protocols, including the one delivered by molecular 
>> probes. I also tried to do the labeling at 4°C to prevent endocytosis but 
>> without result, still the stain was visible as vesicles.  I searched the 
>> list but most discussions about DiI concern neurons or blood vessels. 
>> Does anybody have experience with labeling endothelial cells with DiI? Or 
>> does somebody know of a good non toxic red fluorescent live counterstain 
>> for the nuclear membrane or nucleus, better than DiI?
>> Many thanks in advance,
>>
>>
>> Winnok De Vos (ir.)
>> Academic Assistant
>> ________________________________________________
>>
>> Faculty of bioscience-engineering, UGent
>> Department molecular biotechnology
>> Coupure links 653 (R224)
>> 9000 Gent
>> Belgium
>>
>> tel nr. 09/264.59.71
>> fax nr. 09/264.62.19
>
>
> -- 
> Dr. Andrea J. Elberger
> Professor, Anatomy and Neurobiology
> Director, Confocal Laser Scanning Microscope Facility
> The University of Tennessee, Memphis
> 855 Monroe Avenue
> Memphis, TN  38163  U.S.A.
> tel:    901-448-4101
> FAX:    901-448-7193
> <mailto:[log in to unmask]>
>