Brain tissue preparation for ISH

Technical
1

In the paper “TECHNICAL WHITE PAPER: IN SITU HYBRIDIZATION DATA PRODUCTION” at the “dissection and freezing” section it is mentioned that: “Standard procedures were developed to isolate, cut, fix and pre-treat tissue to preserve macro and cellular morphology and to produce the best signal to noise ratio for ISH”. But at the method’s description it is mentioned that “The brain was rapidly dissected and placed into the OCT.” Does it mean that no prior fixation and/or cryopreservation steps were performed before the brains were frozen?

Thank you for your kindly attention,

Simone Bessa

3

We have reached out to a domain expert who worked on this project and they say that the brains "were not fixed. They were placed onto a piece of foil with pre-chilled OCT on wet ice. Left to acclimate for 2-5 minutes than rapidly frozen in OCT. "