A Cryoprotectant Protocols for Storage of Free Floating Brain Sections - michelletytong/TRGProtocols GitHub Wiki

From http://www.ihcworld.com/_protocols/histology/cryoprotectant.htm

The following protocols are used for long term storage of unstained free-floating brain sections. Transfer brain sections to the cryoprotectant and store at -20°C. Based on purpose of staining, choose the appropriate one for storing brain sections.

Cryoprotectant 1 (Use for Light Microscopy)

To make 500 ml cryoprotectant

Sucrose ---------------------------- 150 g

0.1M PB ---------------------------- 200 ml

Ethylene glycol -------------------- 150 ml Add more 0.1M PB to final volume of 500 ml and store at -20°C. Cryoprotectant 2 (Use for both Electron Microscopy & Light Microscopy)

To make 1000 ml cryoprotectant

Ethylene glycol ------------------- 300 ml

Glycerol --------------------------- 300 ml

0.2M PB --------------------------- 100 ml

Distilled water ------------------- 300 ml

Mix well and store at -20°C.

Cryoprotectant 3 (Use for Immunofluorescence)

To make 1000 ml cryoprotectant

Sucrose ----------------------------- 300 g

Polyvinyl-pyrrolidone (PVP-40) --- 10 g

0.1M PB ----------------------------- 500 ml

Ethylene glycol --------------------- 300 ml Add PVP-40 to 0.1M PB. Stir to dissolve. Slowly add the sucrose to dissolve, and then add the ethylene glycol and bring the final volume to 1000 ml with 0.1M PB. Store at -20°C.