Re: DiD / DiI / DiO versions 'solid' and 'oil'

Posted by Sylvie Le Guyader-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/sorry-tp593651p593653.html

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Steffen

From my work in Zebrafish I'd recommend to purchase DiD solid so you can
dissolve it in what you want. It might be difficult to get it to mix well in
blood if it is diluted in oil.

In live fish we were using DiI and DiO diluted about 5% in Ethanol. You'll
definitely get some dye leaks to neighbouring cells after a few hours so you
will need to image fairly quickly. For fixed fish we were using a saturated
solution of DiI in chloroform. Leaks also occur but it takes several days at
4º.

If you get the solid form, you can try a few things and see what works best.
Powders are kept in a dry box.

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14157 Huddinge
Sweden
[phone]

> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Steffen Dietzel
> Sent: 15 July 2008 19:13
> To: [hidden email]
> Subject: DiD / DiI / DiO versions 'solid' and 'oil'
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello there,
>
> the DiI / DiO related membrane stain DiD from
> Molecular Probes comes in two flavors (as do its relatives):
>
> 'DiD' oil = DiIC18(5) oil (cat# D307) or as 'DiD'
> solid = DiIC18(5) solid (cat# D7757).
>
> Can anyone comment on what the
> advantages/disadvantages of the two flavors are?
>
> The idea for the experiment is to use DiD to
> stain the membranes of endothelial cells of blood
> vessels (from the interior of the vessel), to be
> able to distinguish the endothelial cells better
> from neighboring smooth muscle cells (SMCs)
> Povided that the endothelium is intact, the dye
> should not be able to reach the SMCs - or so ist
> the hope. DiD should be sufficiently in the red
> to allow the simultaneous usage of the green
> channel e.g. for Fluo-4 Ca measurements.
>
> Regards
>
> Steffen
>
> --
>

----------------------------------------------------------------------------
-----------------------

> Steffen Dietzel, PD Dr. rer. nat
> Ludwig-Maximilians-Universität München
> Walter-Brendel-Zentrum für experimentelle Medizin (WBex)
>
> Mail room (for letters etc.):
> Marchioninistr. 15, D-81377 München
>
> Building location and address for courier, parcel services etc:
> Marchioninistr. 27, D-81377 München (Großhadern)
>
> Phone: [phone]
> Fax-to-email:   +49/89/2180-9976509
> skype: steffendietzel
> e-mail: [hidden email]