# Protocol for live-cell imaging of mitochondrial dynamics in adult Drosophila oenocytes

## TL;DR

This paper provides a detailed protocol for live-cell imaging of mitochondrial dynamics in adult fruit fly oenocytes using confocal microscopy.

## Contribution

The study introduces a reproducible method for long-term visualization of mitochondrial morphology in Drosophila oenocytes.

## Key findings

- A protocol for fly preparation, dissection, and mounting for live-cell imaging is described.
- Optimized imaging parameters allow stable visualization of mitochondrial dynamics over time.
- Paraquat-induced mitochondrial fission can be observed using mitoGFP labeling.

## Abstract

Mitochondrial dynamics are essential for cellular homeostasis and can be visualized in adult Drosophila oenocytes using live-cell confocal imaging. Here, we present a protocol for live-cell imaging of mitochondrial dynamics in adult Drosophila oenocytes. We describe steps for fly preparation, dissection of abdominal cuticle to expose oenocytes, and mounting. We then detail procedures for time-lapse acquisition of mitochondria labeled with mitoGFP. Optimized imaging parameters enable reproducible visualization of mitochondrial morphology stably for longer durations.

•Steps for mounting adult Drosophila for live mitochondrial imaging•Procedures for accessing oenocytes through the cuticle for confocal imaging•Instructions for visualizing paraquat-induced mitochondrial fission using mitoGFP•Guidance on quantifying mitochondrial dynamics using Fiji

Steps for mounting adult Drosophila for live mitochondrial imaging

Procedures for accessing oenocytes through the cuticle for confocal imaging

Instructions for visualizing paraquat-induced mitochondrial fission using mitoGFP

Guidance on quantifying mitochondrial dynamics using Fiji

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Mitochondrial dynamics are essential for cellular homeostasis and can be visualized in adult Drosophila oenocytes using live-cell confocal imaging. Here, we present a protocol for live-cell imaging of mitochondrial dynamics in adult Drosophila oenocytes. We describe steps for fly preparation, dissection of abdominal cuticle to expose oenocytes, and mounting. We then detail procedures for time-lapse acquisition of mitochondria labeled with mitoGFP. Optimized imaging parameters enable reproducible visualization of mitochondrial morphology stably for longer durations.