A Septic Pinprick Technique to Introduce Systemic Bacterial Infection in Flies

Samenvatting

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

1. 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.
2. Collect experimental flies 0 - 3 days after eclosion from the pupal case and transfer them onto fresh medium.
3. House the collected flies at a desired temperature (temperatures between 22 °C and 28 °C are generally suitable) until they are aged 5 - 7 days post-eclosion....

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
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