Iniezione di mosche Drosophila adulte: un metodo di consegna composta o etichetta
Published: April 30, 2023
Abstract
Fonte: Nazario-Toole, A. E., Wu, L. P. Assessing the Cellular Immune Response of the Fruit Fly, Drosophila melanogaster, Using an In Vivo Phagocytosis Assay. J. Vis. Exp. (2019).
Questo video descrive come eseguire un’iniezione di mosche adulte vive e include un protocollo di esempio in cui particelle etichettate e blu trypan vengono iniettate negli addome di mosca per saggiare la fagocitosi.
Protocol
Questo protocollo è un estratto da Nazario-Toole e Wu, Assessing the Cellular Immune Response of the Fruit Fly, Drosophila melanogaster, Using an In Vivo Phagocytosis Assay, J. Vis. Exp. 1. Preparare le particelle di fluoresceina per l’iniezione Ricostituire 10 mg di particelle batteriche uccise dal calore disponibili in commercio etichettate con fluoresceina (vedi Tabella dei materiali)ad una concentrazione di 10 mg/mL aggiungendo 990 μL…
Materials
| 5430-10 PicoNozzle Kit | World Precision Instruments | 5430-10 | Holder for 1.0mm pipette |
| E. coli (K-12 Strain) BioParticles, Fluorescein conjugate | Invitrogen | E2861 | Killed E. coli labeled with FITC (Fluorescein). Use to test phagocyte recogntion and uptake of gram-negative bacteria. (~494/~518 nm) |
| Needle Pipette Puller | David Kopf Instruments | Model 725 | |
| Pneumatic PicoPump PV820 | World Precision Instruments | SYS-PV820 | The World Precision Instruments Pneumatic PicoPump PV820 uses differential pressures to hold liquid in the glass needle between injections. The user manually controls short bursts of gas pressure to expel the liquid – allowing delivery of sub-nanoliter volumes. The amount of liquid delivered depends on two main variables – the size of the glass needle opening and the amount of time injection pressure is applied. set the instrument to 100 ms "TIMED" mode. |
| Thin Wall Glass Capillaries | World Precision Instruments | TW100F-3 | Needles for injection. OD = 1.0 mm |
| Trypan Blue Solution (0.4%) | Sigma | T8154 | Used to quench extracellular fluorescence of Fluorescein, Alexa Fluor, or Texas Red labeled particles. |
Cite This Article
Injection of Adult Drosophila Flies: A Method of Compound or Label Delivery. J. Vis. Exp. (Pending Publication), e20113, doi: (2023).