Whole cell dye / membrane dye

[Saar Oz]

Hello everyone,
Can anyone suggest a good dye or approach to visualize a whole cell or the outline of the cell by dyeing the membrane, other than GFP transfection? I will mostly use it to visualize fixed primary neuron cultures.

Preferably something in the blue emission range, so it can be done together with other dyes.

Thanks,

Saar

 

[John Oreopoulos]

Saar,

The dialkylcarbocyanine probes (DiI, DiO, DiD, etc.) work really well  
for staining the cell membrane, thus providing a good outline of the  
cell by fluorescence. Over time these dyes will slowly become  
incorporated into the cell, staining other lipid vesicles. See more  
information here at the Molecular Probes Handbook (no commercial  
interest):

http://www.invitrogen.com/site/us/en/home/References/Molecular-Probes- 
The-Handbook/Probes-for-Lipids-and-Membranes/Dialkylcarbocyanine-and-
Dialkylaminostyryl-Probes.html

I should also mention that these same probes are available from many  
other vendors. Shop around and find a good price.

John Oreopoulos


On 15-Mar-10, at 8:25 AM, Saar Oz wrote:

> Hello everyone,
> Can anyone suggest a good dye or approach to visualize a whole cell  
> or the outline of the cell by dyeing the membrane, other than GFP  
> transfection? I will mostly use it to visualize fixed primary  
> neuron cultures.
> Preferably something in the blue emission range, so it can be done  
> together with other dyes.
>
> Thanks,
>
> Saar
>

[JOEL B. SHEFFIELD]

Depending on your cells, you might also try fluorescent lectins, such
as WGA or ConA.  They don't stain the lipid in the membrane, but
stain associated glycoproteins and glycolipids by virtue of the sugar
groups.  Different cells may respond differently due to cell type
specificity.

Alternatively, there is a set of dyes sold by InVitrogen called FM-4-
64 (that one is red, but there is also a green variety, whose name I
forget).  These incorporate into lipids, and are used with live cells
as tracers for endocytosis.

Best of luck.

Joel

> Saar,
>
> The dialkylcarbocyanine probes (DiI, DiO, DiD, etc.) work really
> well  
> for staining the cell membrane, thus providing a good outline of the
> cell by fluorescence. Over time these dyes will slowly become  
> incorporated into the cell, staining other lipid vesicles. See more
> information here at the Molecular Probes Handbook (no commercial  
> interest):
>
> http://www.invitrogen.com/site/us/en/home/References/Molecular-Probe
> s-
> The-Handbook/Probes-for-Lipids-and-Membranes/Dialkylcarbocyanine-and
> -
> Dialkylaminostyryl-Probes.html
>
> I should also mention that these same probes are available from many
> other vendors. Shop around and find a good price.
>
> John Oreopoulos
>
>
> On 15-Mar-10, at 8:25 AM, Saar Oz wrote:
>
> > Hello everyone,
> > Can anyone suggest a good dye or approach to visualize a whole
> cell  
> > or the outline of the cell by dyeing the membrane, other than GFP
> > transfection? I will mostly use it to visualize fixed primary  
> > neuron cultures.
> > Preferably something in the blue emission range, so it can be done
> > together with other dyes.
> >
> > Thanks,
> >
> > Saar
> >

--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
[hidden email]  
[phone], fax [phone]
http://astro.temple.edu/~jbs

[Sean Speese]

Hi Saar,
   You can also use wheat germ agglutinin (WGA) or Conconavalin-A conjugated to different fluorophores.  I have had good luck staining the with these.  They will also stain intracellular membranes if you permeabilize.

Sean Speese
UMASS Medical School
________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Saar Oz [[hidden email]]
Sent: Monday, March 15, 2010 8:25 AM
To: [hidden email]
Subject: Whole cell dye / membrane dye

Hello everyone,
Can anyone suggest a good dye or approach to visualize a whole cell or the outline of the cell by dyeing the membrane, other than GFP transfection? I will mostly use it to visualize fixed primary neuron cultures.
Preferably something in the blue emission range, so it can be done together with other dyes.

Thanks,

Saar