Protocol for Reversing Clearing of Visikol® HISTO™
In developing Visikol HISTO, our most important goal was to preserve the histological integrity of cleared specimens so that clearing can be reversed. The reversibility of Visikol HISTO means you can validate 3D data against slide-based histopathology
The process of reversing Visikol HISTO is rapid and simple
This protocol is designed to be conducted following clearing and imaging in Visikol® HISTO-1™ or Visikol® HISTO-2™
1. Cleared tissue should be placed directly into a large volume of ethanol (absolute, or histological grade) or methanol. Volume of alcohol should be at least 10-20x tissue volume. Leave tissue at room temperature until opacity has been restored.
2. Larger and more vascular tissues (e.g. whole kidney) may require 2-3 washes of alcohol over the course of several hours.
3. After reversal, samples can be processed directly for paraffin embedding histological preparations.