Domeless Egg Collection and Cell Culture Preparation Day 2

Making New Medium

  • Not enough medium without serum to do the whole process, only about 15mL left and I need 20mL. I decided to make more medium with serum as well just to make everything at the same time (this also saves a 50mL serological pipette)
  • I sort of let the solutions warm up before pipetting, but not completely. The conicals were at room temp by the time I used them on the eggs
  • All steps were in the tissue culture hood
  • 420 medium with 10 % serum
    • 5mL serum
    • 44.5mL 420 medium
    • 500ul 100X antibiotics mix
    • 50ul gentamicin 50mg/mL
  • 420 medium no serum
    • 49.5mL 410 medium
    • 500ul 100X antibiotics
    • 50ul gentamicin 50mg/mL

Collecting Eggs and Cell Culture Prep

  • Followed the exact same protocol as Domeless day 1
  • Instead of setting the flies up again, they were put into the morgue
  • I looked at the flasks after they had settled for the afternoon and they looked good. Lots of single cells, some cell clumps but they were small and few
  • Image is at 20X magnification (plus whatever the camera does)