[Histonet] shrinking glomeruli

[Kim O'Sullivan]

Hi,

Does anyone have any suggestions as to why our mouse glomeruli are
shrinking after coverlipping in DPX? Immunohistochemistry is performed on PLP frozen cryostat sections, and the glomeruli and tubules look fine
upon checking after the chromagen is developed, but once they are
dehydrated, cleared in histosol, and then coverslipped in DPX(also tried
with Vector hardmount) the glomeruli shrink, and the tubules look like
they are spreading out. We have been doing this for years, and not had a
problem. Does anyone know why we would have one now? Could the disease
state of the tissue contribute? Or earlier fixation- could that contribute downstream?

Any suggestions would be appreciated- it is doing my head in!

Kim O'Sullivan
Department of Medicine
Monash Univeristy
Australia