This protocol is available for download on Owens's lab page: https://www.cvrc.virginia.edu/Owens/files/ISH-PLA%20Owens%20lab%20protocol.docx. And this is a published protocol on Nature Methods: https://www.nature.com/articles/nmeth.2332
DAY 1
Step 1: Human or mouse tissue sections
PFA-fixed and paraffin-embedded tissues. Sections: 5µm. (Can try 10um but it is hard to have all important data and all PLA signals focused in the same plane).
Step 2: Immunofluorescent staining
Deparaffinization and rehydration:
- Xylene: 2x5min
- EtOH 100% 2x5min
- EtOH 95% 2x5min
- EtOH 70%: 1x5min
- dH2O: 2x5min
Note: Workstation – tissue tek II.