Cell Culture Preparation of D. innubila Eggs

20220124 Making Yeast Plates

  • Used yeast paste made the week before
  • Then that afternoon, made 2 apple juice yeast plates with 3 very small streaks of yeast on each plate, and these were thin so they would be able to dry well. Also the plates were left for 2-3 nights to dry

20220126 Setting up D. innubila Flies to Lay

  • I tried to set up these flies in a way where I wouldn’t have to knock them out. I took a cage and covered the opening with press n seal, then cut a small V out of one side. Then I used that opening to dump fly vials into
  • When I moved a vial, I covered the hole with a cotton ball
  • This worked for a while, until there were a lot more flies in the cage and every time I took of the cotton ball a bunch would fly out before I could put in the next vial
  • I had to stop adding flies before I had wanted to
  • In the last vial I added, the cotton roll feel out into the cage and squished some flies. I knew I had to take it out so I decided to knock out the flies for as quick as possible to take it out
  • I probably added 10-15 vials to the cage (but this is less than I usually do for melanogaster)

20220127 Day 1 Cell Culture

  • Collected the apple juice plate at ~11am and replaced the cage with a new plate for more laying
  • Picked out all the eggs on the plate: 154 eggs, and put them in a beaker with egg wash before bringing to 4012 to filter and rinse more throuroughly
  • Roughly followed the Prepping Cell Culture Protocol, but also used the modifications from the innubila cell culture last week that did not use trypsin and minimized washes
  • Made fresh 0% and 20% Schneider’s medium with all antibiotics
  • 1 flask was made from these cells
  • Imaged that afternoon:

20220128 Day 1 Cell Culture

  • Collected the apple juice plate at ~10am and spread the flies out over 8 vials to keep in my stocks
  • Picked out all the eggs on the plate: 127 eggs, and put them in a beaker with egg wash before bringing to 4012 to filter and rinse more throuroughly
  • Roughly followed the Prepping Cell Culture Protocol, but also used the modifications from the innubila cell culture last week that did not use trypsin and minimized washes
  • 0% and 20% Schneider’s medium with all antibiotics made yesterday was used
  • 1 flask was made from these cells
  • Imaged that afternoon: