This video demonstrates the visualization of membrane protein trafficking in Drosophila photoreceptor cells using eGFP-tagged TRPL proteins. It details the preparation of transgenic flies for imaging, the capture of fluorescence images of photoreceptor cells, and the quantification of TRPL trafficking by measuring relative eGFP fluorescence in the rhabdomeres.
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Method Article
Studying Membrane Protein Trafficking in Drosophila Photoreceptor Cells
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April 28th, 2025
In This Article
Abstract
Protocol
1. General considerations
- Use Drosophila stocks expressing a permanently rhabdomerally located translocating proteins for analyses regarding protein trafficking (e.g., transient receptor potential-like (TRPL) ion channel:: enhanced green fluorescent protein (eGFP), Arrestin 2 (Arr2)::eGFP).
- Predetermine light exposure conditions of selected flies for the experimental approach.
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Results
Figure 1: Water immersion microscopy workspace. Materials needed are: (A) 15 mL centrifuge.......
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| 15 mL centrifuge tube | Greiner Bio-One | 188271 | |
| Carbon dioxide anaesthesia fly pad | Flystuff | 59-172 | |
| Cold light lamp (KL 1500 LCD) | Zeiss | ||
| Fiji/ImageJ | NIH | ||
| Fluorescence microscope with UV lamp, camera, filter set and software (AxioImager.Z1m, Axiocam 530 mono, 38 HE, ZEN2 blue edition) | Zeiss | ||
| Fluorescent tube | Osram | 4050300518039.00 | |
| Laboratory pipette (20-200 µL) | Eppendorf | ||
| Object slide | Roth | 656.1 | |
| Pipette tips (200 µL) | Labsolute | 7695844 | |
| Plasticine (Blu-Tack) | Bostik | 30811745 | |
| Stereo microscope (SMZ445) | Nikon | ||
| Stereo microscope with UV lamp, camera, filer set and software (MZ16F, MC170 HD, GFP3, LAS 4.12) | Leica |
