Cell Culture Preparation of IMD Null Eggs

20220124 Making Yeast Plates

  • Used yeast paste made the week before
  • Then that afternoon, made 2 apple juice yeast plates with 2 streaks of yeast on each plate

20220125 Setting up IMD null Flies to Lay

  • Set up 8 vials of IMD Null flies to lay on the yeast plate at about 2pm

20220126 Day 1 Cell Culture

  • Collected the apple juice plate at ~9:30am and replaced the cage with a new plate for more laying
  • 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. I have a hypothesis that using the trypsin is hurting the cells because the innubila cells from last week look more alive and larger than all of my other cells
  • Used the 0% FBS and 20% FBS Schneider’s medium with no gentamicin made the week before, and new medium was made for the next day
  • There were a good amount of eggs on the plate
  • Flasks were imaged in the afternoon:

20220127 Day 2 Cell Culture

  • Collected the apple juice plate at ~9:30am and replaced the cage with a new plate for more laying
  • 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.
  • Used the 0% FBS and 20% FBS Schneider’s medium with no gentamicin made the previous day
  • There were so many eggs on the plate, it created a huge pellet
  • Flasks were imaged after settling:

Almost all the circles are cells in these images, it’s pretty incredible!