Home › Cell Biology › The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells
Cell Biology JoVE (Open Access) Citable · DOI

The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells

DOI: 10.3791/50610-v
What you'll learn
  • ✓ Apply Tomato/GFP-FLP/FRT genetic method to create mosaic photoreceptor labeling in Drosophila
  • ✓ Perform live imaging of individual photoreceptor cells over multiple days or weeks
  • ✓ Monitor photoreceptor cell fate changes and degeneration in retinal tissue
Protocol

The Tomato/GFP-FLP/FRT method involves visualizing mosaic photoreceptor cells in living Drosophila. It can be used to follow individual photoreceptor cell fates in the retina for days or weeks. This method is ideal for studies of retinal degeneration and neurodegenerative diseases or photoreceptor cell development.

Difficulty
advanced
Total time
~1–2 weeks per experiment (including fly cross generation, eclosion, and imaging across multiple timepoints)
Model organism
Drosophila melanogaster

Steps

1
Prepare Drosophila for photoreceptor visualization

Set up transgenic fly crosses using Tomato/GFP-FLP/FRT transgenic lines and prepare adult flies for imaging. Allow time for genetic recombination and transgene expression.

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2
Visualize photoreceptor cells in living retina

Mount live adult Drosophila and acquire fluorescent images of GFP and Tomato channels to visualize mosaic photoreceptor labeling patterns in the retina.

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3
Image photoreceptor cells across multiple timepoints

Perform repeated live imaging of the same individual photoreceptor cells over days or weeks to track cell fate changes and developmental or degenerative processes.

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4
Analyze results from high-throughput screening

Quantify and interpret photoreceptor phenotypes from large-scale genetic or chemical screens using the Tomato/GFP-FLP/FRT system to identify genes or conditions affecting photoreceptor biology.

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