Click the back button on your browser window to return to the Weisblat Lab website
Hoechst 33258 Counterstaining
- the Hoechst precipitates with PO4, the embryos must be therefore treated in PO4 free buffer.
- Hoechst stock concentration is 1 mg/ml => must be diluted to the right concentration in any PO4 free buffer (Example: 0.1M Tris, pH 7.5).
- Incubate for +/- 30 min.
- Rinse in the buffer that has been used to prepare the mounting solution (see mounting).
Before observation of any fluorescent dyed embryos, they can be treated with n-propyl gallate, it slows the bleaching and hence increases the delay of observing time. These embryos can also be mounted in 80% (and above) glycerol, the pH is more basic which slows the bleaching.
- Mounting is usually made in 70% glycerol (70% glycerol + 30% buffer)