Tuesday 03/22/16

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.
    • knife was exchanged for a wet knife
    • ordered a new histo cryo (cost here) details here, see p 20.
  • 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.
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