I am working on sections (25um) of adult drosophila brains - specifically their optic lobes. The sections are quite transparent and very difficult to view - so I was thinking of staining them with Eosin red to make them more visible. However, I was wondering if Eosin would interfere with the working of other fluorescent stains such as DAPI and HRP.
My plan was to Golgi stain the brains -> Stain them with Eosin -> Sectioning (25um) -> HRP staining -> DAPI staining -> View under fluorescent microscope
Should I use HRP green instead of red, since Eosin is red as well? Thanks.
I think it is not possible to use eosin with floursent staining
It could be possible to apply DAB staining with theEosin or Cresyl violet
Hello Rathnavel,
If you use HRP method to stain your tissue, you can use Hematoxylin to counterstain nuclei and Eosin to counterstain the rest of tissue.
If you use fluorescence method, you counterstain your nuclei with DAPI or propidium iodide, the rest of your tissue can be seen with endogenous autofluorescence.
It is not necessary to use DAPI counterstaining with HRP method...
If you stain your tissue with eosin before your immunostaining you would have a lot of background (eosin make autofluorescence in green channel (i.e FITC)).
Virginie
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