Fixation
We needed a chemically and mechanically stable tissue to enable multiple rounds of labeling. At the same time, we wanted to be able to clear and label the tissue afterwards for intact tissue imaging. So after testing out different fixatives, we developed an optimized fixation solution (4% paraformaldehyde [PFA], 1% glutaraldehyde [GA]) that best preserved antigenicity and provided chemical and mechanical stability.
SWITCH worked on 86/90 (96%) antibodies that we tested, while CLARITY worked on 53/66 (80%) and PFA on 83/90 (92%).
