A modified cleansing procedure to obtain large freeze-fracture replicas

J Microsc. 1985 Feb;137(Pt 2):185-8. doi: 10.1111/j.1365-2818.1985.tb02576.x.

Abstract

A method for cleaning freeze-fracture replicas is elaborated which avoids their rolling up and fragmentating during successive steps. Immediately after freeze-fracturing, the replicated tissue is slowly thawed from 77 K to room temperature on solid methanol. Safe passage through the subsequent cleaning steps is facilitated by using solutions of gradually increasing surface tension. After at least 24 h of fixation in methanol, the tissue is digested in half-strength bleach containing 5% ethanol. Following further cleaning in full-strength bleach, the remaining organic material attached to the replica is dissolved overnight at room temperature in 50% saturated sodium hydroxide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Freeze Fracturing*
  • Histological Techniques*
  • Liver / ultrastructure*
  • Methanol
  • Myocardium / ultrastructure*
  • Rats
  • Sodium Hydroxide

Substances

  • Sodium Hydroxide
  • Methanol