Chapter 49 - Preparation of Organotypic Hippocampal Slice Cultures

Abstract

This chapter provides a concise description of the steps involved in preparing and maintaining organotypic hippocampal slice cultures. There is variability in the amount and type of anticoagulants contained in commercial plasmas. Fill the dish with enough 95% ethanol to cover the coverslips and soak them overnight. Replace the 95% ethanol with 100% ethanol and soak overnight. Sterilize instruments by dunking in 70% ethanol and then in 95% ethanol. Large scissors and spatulas should be flamed in the alcohol lamp after removing from 95% ethanol. It is recommended that you start with rat or mouse pups that are 5–7 days old. Younger animals can also be used, but the dissection will be more challenging. Animals older than 10 days rarely survive more than a few days in vitro. Hold top of head nose down over the petri dish. Using a spatula dipped in HBSS, push the brain stem, cerebellum, and midbrain down gently, leaving the cortex and hippocampus inside the dorsal skull.