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Related Experiment Video

Updated: Nov 1, 2025

Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy

Array Tomography Workflow for the Targeted Acquisition of Volume Information using Scanning Electron Microscopy

Published on: July 15, 2021

High-precision targeting workflow for volume electron microscopy.

Paolo Ronchi¹, Giulia Mizzon¹, Pedro Machado¹

¹Electron Microscopy Core Facility, European Molecular Biology Laboratory, Heidelberg, Germany.

The Journal of Cell Biology

|June 23, 2021

Summary

This summary is machine-generated.

This study presents a new workflow for precisely targeting fluorescently labeled cells for high-resolution volume electron microscopy (vEM) using focused ion beam scanning electron microscopy (FIB-SEM). This method enables detailed ultrastructural analysis of specific cells within complex biological samples.

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Last Updated: Nov 1, 2025

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Area of Science:

Cell Biology
Microscopy Techniques
Structural Biology

Background:

Accurate interpretation of cellular ultrastructure often requires 3D volume electron microscopy (vEM).
Scanning electron microscopy (SEM) methods like focused ion beam SEM (FIB-SEM) are valuable for vEM, but precise targeting of specific cellular volumes within large samples is difficult.
Existing methods lack the precision needed to target specific cells or subcellular structures for detailed vEM analysis.

Purpose of the Study:

To develop and validate a workflow for precise targeting of fluorescently labeled cells and subcellular structures for FIB-SEM acquisition.
To enable high-resolution ultrastructural analysis of specific cellular components within complex biological tissues.
To overcome the limitations of current vEM targeting strategies.

Main Methods:

A workflow combining fluorescence preservation during sample preparation with confocal microscopy for guiding resin block trimming.
Utilizing confocal maps of fluorescence signals to precisely locate target volumes within the resin block.
Employing laser branding on the resin block surface to create accurate landmarks for FIB-SEM positioning.

Main Results:

Successful precise targeting of single cells within large biological samples, including mouse mammary gland organoids, Drosophila tracheal terminal cells, and Drosophila ovarian follicular cells.
Acquisition of high-resolution vEM data from specifically targeted cellular volumes.
Discovery of previously unappreciated ultrastructural details in the analyzed samples.

Conclusions:

The developed workflow allows for micrometer-precision targeting of fluorescently labeled cells for FIB-SEM analysis.
This method significantly enhances the ability to perform detailed ultrastructural investigations on specific cellular components within complex biological contexts.
The approach opens new avenues for exploring cellular ultrastructure with unprecedented detail.