Infecting Flies with Bacterial Suspension Using a Nanoinjector
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 the nano-injection of bacterial suspension into the fly body cavity, resulting in bacterial multiplication and infection. The fly's immune response clears some bacteria, while others escape immune clearance and survive within the host causing an infection.
Protocol
1. Collect and Prepare Flies Rear Drosophila 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 me…
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 | |||
| Microscope | Olympus Corporation | SZ51 | |
| Drosophila Vials polystyrene | VWR international | 89092-720 | |
| Nosterile Large Cotton Balls | Fisher brand | 22-456-883 | |
| 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 | |
| Vertical Capillary Puller | Kopf Needle Pipette Puller | ||
| 3.5'' Replacement glass Capillaries for Nanojet II | Drummond Sientific Company | 3-000-203-G/X | |
| Nanoject II | Drummond Sientific Company | 3-000-204 | |
| Forceps | Fine Science Tools | 11255-20 | |
| 10mL Syringe | BD | 309604 | |
| Mineral Oil, White, light | Macron Fine Chemicals | 6358-10 | |
| 1.5mL Microcentrifuge tubes; Seal Rite | USA Scientific Inc. | 1615-5500 |
Tags
Citer Cet Article
Infecting Flies with Bacterial Suspension Using a Nanoinjector. J. Vis. Exp. (Pending Publication), e21649, doi: (2023).