Mon-Fri 9:00-5:00
Cryoprotectant Protocols for Storage of Free-Floating Brain Sections
Home » Protocols  »  General Histology Protocols  »  Cryoprotectant Protocols for Storage of Free-Floating Brain Sections
Cryoprotectant Protocols for Storage of Free-Floating Brain Sections

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.

Leave a Reply