Going Through the Cell Culture Flasks Since I Started D. innubila and No Longer Using Trypsin

innubila

01/27

  • Has not been fluid changed yet
  • The plates I used for laying were the large apple juice plates, so I got few eggs
  • However, I do see some large cells in the flask, and they look like they may be replicating!
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics I’ll want to talk to Kent about that first image. He showed me on Wednesday the 01/14 flask that he infected with DiNV and was thinking that you could see infection in some cells. They looked sort of like the first image here so maybe it’s not CPE.

01/28

  • Has not been fluid changed yet
  • Didn’t have many eggs (like 01/27)
  • Flasks have no visible cells, although I did not look exhaustibly
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics

01/13

  • Was fluid changed (3mL removed and replaced) on 02/10
  • Lots of cell growth! Lots of different types of cells growing, some really huge, some long and filimenty, some small round ones. I think a lot of the big ones are dead, some are not attached anymore. But they don’t necessarily have to be attached to be alive still.
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics

Kent took this flask on 02/18 and removed 2mL and put in a new flask, and added 5mL new medium. This was probably 10% FBS

02/03

  • 2 flasks, has not been fluid changed
  • Not a lot of cells, but some and they look alive. These are worth keeping, and it might be a good idea to fluid change. I have a video of a clump of cells contracting
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics

02/04

  • 2 flasks, has not been fluid changed
  • Not a lot of cells, similar to 02/03
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics

02/18

  • 1 flask, has not been fluid changed
  • Some cells, there were not a lot of eggs. Also the plate had sat with the flies for an extra day bc of snow day
  • Medium used: 10% FBS Schneider’s medium w/all antibiotics

melanogaster

SPZ
01/20

  • Has not been fluid exchanged
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics
  • Not many, if any, visibly alive cells
  • Did not take picture, looked like 01/28 innubila flasks 01/21
  • Same as above 01/20

IMD Null
01/26

  • 2 flasks, was fluid changed (3mL removed and replaced) on 02/10
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics
  • Cells are smaller than those nice huge innubila ones, but I think there are clumps that are still alive and might be growing. These are worth keeping 01/27
  • 2 flasks, was fluid changed (3mL removed and replaced) on 02/10
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics
  • I started out with a TON of eggs, I should have made 3 or 4 flasks instead of 2
  • These are super overwhelmed with what I think is debirs/cellular junk. I don’t think it’s contamination but it could be. Either way, I don’t think anything will come of these flasks

Myd88
02/02

  • 2 flasks, was fluid changed (3mL removed and replaced) on 02/10
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics
  • There are some cells and some clumps, hard to tell if much growth or not, but these might be doing ok 02/03
  • 3 flasks, was fluid changed (3mL removed and replaced) on 02/10
  • Medium used: 20% FBS Schneider’s medium w/all antibiotics
  • Lots of alive and large cells! These actually look big like the innubila ones. It does look like they might be dividing too!

Dicer2
02/10

  • 2 flasks, no fluid change
  • Medium used: 10% FBS Schneider’s medium w/all antibiotics
  • Lots of debris but some cells too that look good, and might be dividing

02/11

  • 3 flasks, no fluid change
  • Medium used: 10% FBS Schneider’s medium w/all antibiotics
  • Also lots of debris, but still might be ok. Didn’t see any nice sections of the flask with a lot of cells but I didn’t look exhaustively