Home › Cell Biology › Live Imaging of GFP-labeled Proteins in Drosophila Oocytes
Cell Biology JoVE (Open Access) Citable · DOI

Live Imaging of GFP-labeled Proteins in Drosophila Oocytes

DOI: 10.3791/50044-v
What you'll learn
  • ✓ Prepare Drosophila flies and dissect oocytes for fluorescence microscopy
  • ✓ Mount and position oocytes on microscope slides for live imaging
  • ✓ Acquire high-quality live-cell images of GFP-tagged proteins in oocytes
  • ✓ Identify autofluorescent and GFP-labeled structures in real-time imaging data
Protocol

A protocol for live imaging of GFP-tagged proteins or autofluorescent structures in individual Drosophila oocytes is described.

Difficulty
intermediate
Total time
~45 min to 1 hour per fly preparation and imaging session
Model organism
Drosophila melanogaster

Steps

1
Prepare Drosophila flies for dissection

Select and prepare transgenic flies carrying GFP-tagged protein constructs. Ensure flies are at appropriate developmental stage and anesthetized or immobilized for safe handling.

▶ 01:28
2
Dissect and prepare oocytes for imaging

Extract ovaries from flies and isolate individual oocytes. Mount oocytes on microscope slides with appropriate mounting medium to preserve fluorescence and allow optical access.

▶ 02:15
3
Acquire live-cell fluorescence images

Position oocyte sample on an upright compound microscope equipped with fluorescence optics. Capture time-lapse or single-frame images of GFP and autofluorescent signals while minimizing photobleaching.

▶ 04:33
4
Analyze autofluorescent and GFP structures

Review acquired images to identify and characterize GFP-tagged protein localization and autofluorescent cellular structures. Document morphology and subcellular distribution patterns observed in live oocytes.

▶ 06:18
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