- Anesthetic (ketamine 50 mg/Kg and xylazine 4.5
mg/Kg – varies according to institutional protocols)
- Peristaltic pump (e.g.,
Gilson) with tubing and connectors
- Dissection tools
(scissors, fine scissors, spring scissors, tweezers, spatula, according to
preferences)
- PFA stock solution (e.g., 16% PFA solution, Electron Microscopy)
- pH-meter and related
reagents/tools
- Perfusion needle
(preferred: 27 gauge)
- Small plastic bags
for storage of brains in -80°C freezer
- Solid PFA waste
collection bin
- Liquid PFA waste
collection bin
Tissue processing and sectioning
-Tissue freezing medium (e.g., Tissue-Tek O.C.T. Compound, Sakura)
-Cryostat (e.g., Leica CM3050S with accessories)
- Small amount of dry ice
-Antifreeze solution 1l (300 ml Glycerol, 300 ml Ethylenglycol, 400 ml dest. Water, 5.18 g Na2HPO4, 1.57 g NaH2PO4;
-30% Sucrose solution 100 ml (100 ml 0.1 M PB, 30 g Sucrose)
-Cryotubes (e.g., CryoPure tubes, Sarstedt)
Immunofluorescence
staining
-Pre-cut brain sections (30-50 µm thick)
-Normal donkey serum (S30-100ML, Sigma-Aldrich)
-12-well-plates with netwell inserts (e.g. Corning Costar Netwell)
-Hard-drying mounting medium (recommended ProLong Diamond, ThermoFisher Scientific)
-Designated container for storage of microscopy slides
- Water bath with integrated shaker, temperature adjustable (e.g. GFL-1083)
- Orbital shaker (e.g. Heidolph Duomax 1030)
-Platform shaker (e.g. Heidolph Unimax 1010)
-Optional: DAPI, Sigma-Aldrich, D9542-5MG, 1:10,000 of 5 mg/ml