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.

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