Measuring E. coli Bacterial Load in Drosophila melanogaster following E. coli Infection
Published: September 29, 2023
Abstract
Source: Khalil, S., et al. Systemic Bacterial Infection and Immune Defense Phenotypes in Drosophila Melanogaster. J. Vis. Exp. (2015)
In this video, we demonstrate a technique for measuring the E. coli bacterial load after injecting E. coli into the thorax of Drosophila melanogaster.
Protocol
1. Assay Bacterial Load Post-infection Measure bacterial load. At a prescribed time post-injection, place each fly into a microcentrifuge tube on ice. Add 250 µl PBS to each tube and homogenize the flies either using a pestle or a bead beater. Plate the homogenate on an LB agar plate, either using a spiral plater or a serial dilution. To plate using serial dilution, transfer each fly homogenate to the first row of a 96-well plate. <…
Divulgazioni
The authors have nothing to disclose.
Materials
| 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 | |
| 1.5mL Microcentrifuge tubes; Seal Rite | USA Scientific Inc. | 1615-5500 | |
| Motorized Pestle; Talboys Laboratory Stirrer | Troemner | 103 | |
| Talboys High Throughput Homogenizer | OPS Diagnostics | 930145 | |
| 5/32'' Grinding Balls | OPS Diagnostics | GBSS 156-5000-01 | |
| Vortex Genie | Scientific indurstries inc. | G560 | |
| Multichannel Pipettor (10uL-300uL) | Sartorius | 730360 | |
| WASP2 Whitley Automated Spiral Plater | Microbiology International | ||
| ProtoCOL automated colony counter / plate counter/ plate reader | Microbiology International | ||
| Incubator | Powers Scientific, Inc | DROS52SD |
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Citazione di questo articolo
Measuring E. coli Bacterial Load in Drosophila melanogaster following E. coli Infection. J. Vis. Exp. (Pending Publication), e21651, doi: (2023).