This is the result of immunohistochemical staining using paraffin-embedded mouse sections. Dewaxing, hydration, antigen activation (ph6.0 citrate, autoclave), blocking, and incubation of primary and secondary antibodies were experienced. DAB used for color development and counterstaining was also performed. The mounting medium was used with softmount.
The results showed artificial disturbances in the figure, like bubbles or crystals. Does anyone know what is causing this? What should be done to avoid it?
before mounting sections you should wash your lam with passaging through very strong base like KOH and then put them in D-water and next strong acid like HCL and again DW in this way your lam will became very clean then instantly put them in fresh filtered gelatin solution and leave them dry for a night and make sure not became polluted by covering them.
after sectioning and dewaxing, passage sections in another bath and make sure sections perfectly dewaxed when mount on lam also surface of lam most not encounter with wax specially under of mounted section.
use sampler for Entellan dropping on lam and make sure on bubble remain under lamella by pushing lamella.
do not heat Entellan or gelatin coated lam because distribution became disrupted and making bubble.
good luck.