Testing Fluids From Primary innubila Cells Infected with DiNV From Kent by Infecting D. innubila Flies with the Fluid
Goal is to test the infectability of this fluid on adult flies. The sample is from day 26 after DiNV infection on primary innubila cells.
Flies were aged to be 7-10 days old for the infection. Flies emerged 9/5-9/8 and were infected on the 15th. I also infected both the males and the females for this experiment. Additionally, for this experiment I took day 0 flies and froze them. These are flies that are poked (either cell culture medium or DiNV) and then frozen immediately. The control for this experiment is Schneider’s medium with 10% FBS and antibiotics (Streptomycin, Penicillin, Gentamicin, and Amphotericin B). This is the cell culture medium the primary cells were in at the time, however it is not from an actual tissue culture flask, so there are no cell components in the liquid.
The goal was to infect ~40 males for the experiment, 20 males for control, and as many females as I had. I had not planned on infecting females when I set up the vials to age so I only had a few in each vial to keep the males happy. Vials were set up to have ~12 males and ~5 females each.
The infection process followed this general protocol except that it was virus and not bacteria, and that the flies will be monitored for 14 days, and I will freeze the dead flies every 3 days. The virus was thawed from -80 on ice, and then I pipetted ~40ul into the top lid of a 0.6mL tube and would dip the needle in there for infections. I did the same with the medium.
Vials and their contents:
| vial | treatment | time | sex | n# |
|---|---|---|---|---|
| 1 | medium | 2:57 | M | 11 |
| 2 | medium | 2:57 | F | 5 |
| 3 | DiNV | 3:09 | M | 7 |
| 4 | DiNV | 3:09 | F | 5 |
| 5 | DiNV | 3:16 | F | 6 |
| 6 | DiNV | 3:16 | M | 4 |
| 7 | DiNV | 3:20 | M | 10 |
| 8 | DiNV | 3:20 | F | 4 |
| 9 | medium | 3:36 | M | 10 |
| 10 | medium | 3:36 | F | 4 |
| 11 | DiNV | 3:47 | M | 9 |
| 12 | DiNV | 3:47 | F | 6 |
| 13 | DiNV | 3:55 | M | 9 |
| 14 | DiNV | 3:55 | F | 7 |
| 15 | DiNV | 4:02 | M | 12 |
| 16 | DiNV | 4:02 | F | 7 |
Flies that were frozen day 0:
| tube # | sex | treatment |
|---|---|---|
| 1 | M | medium |
| 2 | F | medium |
| 3 | M | DiNV |
| 4 | F | DiNV |
| 5 | M | DiNV |
| 6 | F | DiNV |
| 7 | M | medium |
| 8 | M | DiNV |