Staining of neurons: biocytine vs. lucifer yellow?

SJM Guzman jose.guzman at medizin.uni-leipzig.de
Thu May 12 10:17:07 EST 2005


Hi Thomas:

It actually depends more on your equipment. Biocytine labeling is 
suitable for light microscopy whereas Lucifer yellow implies the use a 
fluorescence microscope.

Biocytine labeling is a much more older (and classic) method, which 
demands fixing overnight, developing with peroxidase, etc.. whose 
conditions are well-described in the literature,and don't vary too much 
for different cell types. The dendritic arbourization is easily visible 
through the staining, and provides a good morphology description.

Lucifer yellow does not need to be developed, and has the advantage of 
the plausible use of the high-resolution fluorescence techniques 
(confocal, two-photon...) and posterior digitalization and imaging. 
Moreover, Lucifer yellow is essential if you want to label your tissue 
with fluorescence antibodies, to describe certain protein localization 
(i.e somatic vs. dendritic) . However, I find the morphology is not that 
clear as this showed by Biocytine, unless you use confocal-microscopy.

Depending on the tissue and on your experience you can get a good by 
both staining methods. I saw wonderful 3D pictures reconstruction with 
Lucifer yellow, but I have to admit that Biocytine labeling of the 
dendritic arbourization is better.

For Lucifer yellow you should work in darkness, to avoid the fluorophore 
bleaching.

I guess you're filling the cells with patch-micropipettes. For both 
methods, you should take care of forming the outside-out configuration 
after filling the cell (10-20 min), to avoid the diffusion of your 
marker into the slice (this is specially unpleasant with Biocitin). 
Biocitin solution concentrations are in the range of 1-2% and Lucifer 
yellow is about 0.6%.

I checked you're writing from Germany... I'm electrophysiologist working 
for the medical faculty of the University of Leipzig, we have some 
experience here for such labeling (also in cortex), so if you have any 
question, please contact me!

Viele Grüsse!

SJM Guzman



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