9:00 am – 5:00 pm
- Fill LN2 (upstairs dewar is empty need to go downstairs to fill)
- Poly-lysine coat newly cleaned 50 mm coverslips — dipped in pure poly-D lysine solution
- substantial residue after air drying. The adhered samples seem to resist washing
- dip into ddH2O to dissolve excess salts. Hopefully the samples still stick.
- 10:20 am – started cool down of LN2
- cryo-knife has dissappeared.
- cutting 1.5 um sections with Diamond cryotrim knife
- settings: -51C knife, -50C sample, -50C chamber.
- most of the morning cutting was very smooth, readily get 10 sections
- other parts in need of replacing
- both perfect loops are damaged
- all the eyelash brushes are decapitated. Made 2 new ones.
sample prep for DNA staining
- ran staining protocol for two large coverslips
- samples now in 50% formamide 2x SSC in 4C, ready for staining
- actually could have had samples ready for tomorrow, too bad I already gave away my scope time.
- should label with libraries L8-cy3 and L7-cy3 tomorrow.