A Septic Pinprick Technique to Introduce Systemic Bacterial Infection in Flies
Published: September 29, 2023
Abstract
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.
Protocol
1. Collect and Prepare Flies Rear D. melanogaster under the desired experimental conditions. Take care not to overcrowd the flies during rearing and make sure that larval densities are consistent across treatments since environmental conditions during the larval stage can profoundly affect immune defense phenotypes during the adult stage. Collect experimental flies 0 – 3 days after eclosion from the pupal case and transfer them onto fresh medium. House the collect…
Divulgations
The authors have nothing to disclose.
Materials
| 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 |
Tags
Citer Cet Article
A Septic Pinprick Technique to Introduce Systemic Bacterial Infection in Flies. J. Vis. Exp. (Pending Publication), e21650, doi: (2023).