Source: Khalil, S., et al. Systemic Bacterial Infection and Immune Defense Phenotypes in Drosophila Melanogaster. J. Vis. Exp. (2015)
This video demonstrates a technique to infect Drosophila by introducing a pathogenic bacteria directly into their body cavity. This method bypasses epithelial barriers and other passive defense mechanisms, allowing for the induction of systemic infection. Upon infection, the resulting immune response is assessed by monitoring fly mortality and performing downstream assays.
1. Collect and Prepare Flies
2. Culture and Prepare Bacteria
3. Infect the Flies
NOTE: As Drosophila immunity is influenced by circadian rhythm, it is important to perform infections at a similar time of day across experimental replicates.
The authors have nothing to disclose.
Incubator | Powers Scientific, Inc | DROS52SD | |
Paintbrush | |||
CO2 Flypads | FlyStuff | 59-114 | |
CO2 | Airgas | CD FG50 | |
Drosophila rearing mix | |||
6 oz Square Bottom Bottles, polypropylene | Genesee Scientific | 32-130 | |
Nosterile Extra Large Cotton Balls | Fisher brand | 22-456-882 | |
Microscope | Olympus Corporation | SZ51 | |
Drosophila Vials polystyrene | VWR international | 89092-720 | |
Nosterile Large Cotton Balls | Fisher brand | 22-456-883 | |
2L flask | VWR international | 89000-370 | |
Petri Dishes with Clear Lids, Raised Ridge; 100 . x 15 mm; | VWR international | 25384-302 | |
LB Agar, Miller | Difco | 244520 | |
Inoculating Loop | VWR international | 80094-488 | |
Rainin Clasic Pipettes in various sizes 0.1 µl to 2 µl, 2 µl to 20 µl, 20 µl to 200 µl, 100 µl to 1000 |
Rainin | PR-2 PR-20 PR-200 PR-1000 |
|
Micropipette tips (assorted sizes) | VWR international | 30128-376 53503-810 16466-008 |
|
Luria Broth Base, Miller | Difco | 241420 | |
Disposable Culture Tubes Borosilicate Glass | VWR international | 47729-576 | |
S-500 Orbital Shaker | VWR international | 14005-830 | |
Centrifuge | VWR international | 37001-300 | |
PBS pH 7.4 10x | Invitrogen | 70011044 | |
SmartSpec 3000 Spectrophotometer | Bio-Rad | 170-2501 | |
Semimicrovolume Cuvettes | Bio-Rad | 223-9955 | |
Forceps | Fine Science Tools | 11255-20 | |
Mineral Oil, White, light | Macron Fine Chemicals | 6358-10 | |
Minutein pins | Fine Science Tools | 26002-10 | |
1.5mL Microcentrifuge tubes; Seal Rite | USA Scientific Inc. | 1615-5500 |