11:15 A -7:30P, 9:30P – 11:45 P
UltraCryo coverslip DFISH
- Hot washes for DFISH coverslips
- RT wash
- check sections on coverglass A. Be very careful to keep sections covered with buffer.
- Staining definetely works. Signal somewhat dimer than expected, but about 1 hr at 55C is probably too agressive a hot wash for 25mer of only As and Ts is substantially above Tm for the probe.
- Some sections very wavy, not at all flat. Need to get better at this sectioning. For those that are flat though, they should be much thinner and closer to the coverglass than the cells.
- Some very nice flat sections.
UltraCryo coverslip AB
- Rinse coverslip in coplin jars
- Green humidity chamber– supports not flat enough — slide is angled and corner of slide dries out.
Imaging on STORM2
- Run log here
- STORM of cells with H3K9me2
- STORM of embryo sections
- Fairly certain many more of the sections were flat to start with.