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Thank you Katrin! We'll look into that.

Best,

Claude
________________________________________
Dr. Claude Messier 
School of Psychology / Ecole de psychologie 
University of Ottawa / Université d'Ottawa 
136 Jean-Jacques Lussier Room 2076A
Ottawa Ontario
K1N 6N5
(613)562-5800 ext 4562
FAX: (613) 562-5147

Le 2013-05-28 à 08:480, Katrin roth <[log in to unmask]> a écrit :

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> 
> Dear Claude,
> 
> we also used EdU click-it system. To minimalize the EdU usage we used 50ml Falcons in the drinking bottles and we generated little agarose pads with the EdU which is feeded.
> 
> All the best
> Katrin
> ________________________________________
> Von: Confocal Microscopy List [[log in to unmask]] im Auftrag von Claude Messier [[log in to unmask]]
> Gesendet: Dienstag, 28. Mai 2013 14:31
> An: [log in to unmask]
> Betreff: Imaging GFP with BrdU
> 
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> 
> Although this is not a confocal question proper, I would appreciate
> suggestions for a immunohistochemistry problem. We are trying to visualize
> GFP in the brain of transgenic mice expressing GFP in oligodendrocyte
> precursor cells. In order to identify dividing oligodendrocyte precursor cells,
> we also want to use BrdU labelling. The problem is that the acid treatment
> that is needed to obtain BrdU staining also abolish the GFP visualization. GFP
> staining is not extremely strong to start with so we have used anti-GFP
> secondaries to improve visualization. We have manipulated several variables
> associated with the acid treatment (length, concentration, heat, pH) or
> fixation (mostly length of fixation) to no avail. This is a common problem
> with BrdU since the acid treatment interferes with some antibodies. Although
> we have used the EdU click-it system successfully, we are testing a large
> number of animals and the cost of using EdU is significant if not prohibitive.
> So, any suggestion to obtain BrdU staining while preserving endogenous GFP
> stain?
> Many thanks!
> Claude