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This fixative is designed as a substitute for Formalin and can be used in your normal protocol, with the following application notes to be considered:
Fixation Times | |
Cells | 15-20 minutes |
Sections of Slides | 45 Minutes |
1cm3 | 2-3 hours |
Clearing Solution 100 is a non-crosslinking fixative. Tissue will be softer and will have a more natural appearance than Formalin-fixed tissues. If firmer tissue is desired, use four parts Clearing Solution 100 to one part ethanol.
Do not predigest!!! If your current protocol contains a predigestion step (using Proteinase K, trypsin, etc.), this step must be omitted. Clearing Solution 100 does not crosslink tissue; therefore, no predigestion is required.
After perfecting your in situ hybridization or immunohistochemistry procedure using Clearing Solution 100, you may find that you can decrease your primary probe or primary antibody concentration by ½ to 1/10% and obtain the same or better results.
The osmolality of 1:10 dilution of this fixative in diH2O is approximately 340-360 mOsm/kg H2O. Adjustments will need to be made with sodium chloride or sucrose if working with marine (saltwater) organisms.