Test Infections: Infecting Myd88 and W118 Flies with DiNV and Learning How to Infect Flies

Set Up

  • Clean C02 pad and area with 95% ethanol
  • Clean needle, feather brush, and poker/probe with 95% ethanol
  • Virus sample is: Day 15 DiNV 3-22
  • Thawed virus sample on ice
  • Pipette virus liquid into a “free” cap of a 1.5mL tube to easy access
  • Had ice bucket ready
  • Made up 3 tubes per fly line and per sex that said the line, sex, and “day 0” for immediately freezing some flies

Process

  • Knocked out flies on pad (separate flies by sex if not done so already)
  • Maneuver 1 fly to an open space on the pad and position on their side so you can see under the wing (works best for me if the underwing is facing the right hand)
  • Dip needle in virus and poke the fly under the wing, where it looks like they have thinner exoskeleton
    • I did not always poke them in the same place
  • Poke through not quite the whole taper of the point of the needle
    • This was not always done correctly too
  • The needle should be able to pick up the fly, so you can move it to the other side of the C02 pad and use the probe to gently slide it off
  • For the first 3 flies of each sex and line, I put them in individual tubes for freezing as day 0 samples
  • The rest of the flies got put in fresh food vials split up by sex and line and let sit on their side for a long time to recover and wake up
    • Flies that have been stabbed take a longer time to recover after C02
  • Between sexes and lines of flies, dip the probe and needle in 95% ethanol and allow to dry
  • Afterwards, clean the probe and the brush with ethanol
  • Dispose of the needle in the sharps
  • Clean the CO2 pad with ethanol and let bubble off
  • 18 W1118 males were infected, 14 put in the food vial
  • 20 W1118 females were infected, 17 put in the food vial
  • 19 Myd88 males were infected, 16 put in the food vial
  • 24 Myd88 females were infected, 21 put in the food vial

20220505

  • 3 days post infection
  • So far no mortality for any vial
  • Put flies on CO2 pad and individually froze half of each set of flies for day 3 samples
  • 7 W1118 males frozen
  • 8 W1118 females frozen
  • 8 Myd88 males frozen
  • 10 Myd88 females frozen
  • Other flies were left to go for another 3 days