40X coronal stomach section, stained for actin. Click for high res. Scale bar = 200um.
Frozen section, wash off excess OCT in RT dH2O, 10min permeabilization in 100% acetone, 30min block in Toaster's Übermegablocking Buffer*, added primary polyclonal Ab without washing, incubate 1h @ RT, wash 3X in Washy Wash Buffer**, add labeled actin at 1:200 and secondary labeled Ab against primary at 1:10, incubate 1h @ RT, wash 3X in Washy Wash Buffer, tap off all excess wet, 2 drops Prolong Gold Antifade + DAPI (@RT), coverslip w/out bubbles, cure O/N in a dark box.
The resolution I managed to get here is so exquisite and the result so beautiful that I am very tempted to get a 5'x6' giclee print of it and hang it on my everything.
*Toaster's Megablocking Buffer: 1% BSA, 1% FBS, 1% NGS in 1X PBS; to make Toaster's Übermegablocking Buffer add 1:10 CD16/CD32 to block endogenous Fc antibody receptors in the tissue. **Washy Wash Buffer: 2% FBS, 0.02% NaN2 in 1X PBS