For nuclei staining, i washed my DAB stain slides with either PBS/dH2O and add few drops of methyl green, but it did not work even if i stay for 60 mints. If somehow some slight counter stain is there, it washes away with ethanol during mounting procedure.
I have attached a ppt file, JPEG photo stained with a cytoplasmic antibody VEGF/BDNF/ NR2a. I am confused, whether the arrow stained is my cytoplasmic targeted protein or these rounded cell indicates microglia or some inflammatory cells just like we find same cells in nissl/HE staining.
Your answers will be greatly appreciated.
Regards
Katja Berger, i just dip the slides in graded ethanol ( 3-4 seconds each).
Try to use some DAB reaction enhancers like Co and/or Ni (or both together) ions incorporated directly to DAB developing solution. There are two advantages of this: 1/ more intense DAB staining results, 2/ since these ions shift the DAB color to dark or black you can more effectives discriminate the false/positive DAB signal/results.
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