High-Resolution Confocal Imaging of Pathogen-Infected Drosophila Hemocytes

Place a slide containing fixed hemocytes onto the confocal microscope stage.

These hemocytes are obtained from Drosophila melanogaster larvae and have been infected with a bacterial pathogen.

The hemocytes are labeled with green fluorescent protein, their nuclei are stained with a blue dye, and the bacteria express red fluorescent protein for visualization.

Configure the microscope to detect all three fluorophores to track cellular compartments and pathogens simultaneously.

Using a high-resolution objective, focus on the target hemocyte to capture subcellular details.

Adjust the laser power and detector gains to achieve appropriate sample exposure and optimize signal clarity.

Scan multiple Z-planes to ensure the exposure level is appropriate.

Set the top and bottom Z positions to image the entire hemocyte.

Use scanning zoom to isolate the hemocyte of interest and acquire a high-resolution 3D image.

This method enables visualization of pathogen localization within the hemocyte.

Configure the confocal microscope for three-color imaging of DAPI, EGFP, and mCherry.

Place the sample on the microscope and focus on the sample using a 63X/1.4 numerical aperture objective. Locate desired hemocytes in