Unfixed snap- frozen spleen.

Cryostat section ( 10 microns thick/thin) was mounted onto a superfrost Plus slide.

Dried overnight at RT, in an operating fume hood ( I always air-dry cryosections under an airflow O/N. Additionally, for Formalin-fixed tissues that are then frozen, dry in a 50 C oven to minimise loss of sections…imho)

Then, fixed in 10% Formalin in PBS for 15 minutes.

Rinsed in dw x2, rinsed in TBS x2 before ringing section using a hydrophobic pen ( wonderful invention…deserves to be recognised)

Whatever happened to the Israeli hydrophobic pen?

Is it still available?

I recall requesting  then using a free pen.

Excellent!

Primary Ab dilution factor of 1/500, using indirect IF ( Alexa 594 and Hoechst at 1/20K).

FFPWS needs, in my lab,a lower dilution factor ( 1/100)

The submitted image is slightly “fuzzy”. I think that this is due to immersion oil on the objective lens.