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

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Published on: July 15, 2021

Array tomography: imaging stained arrays.

Kristina D Micheva, Nancy O'Rourke, Brad Busse

Cold Spring Harbor Protocols

|November 3, 2010

Summary

This summary is machine-generated.

Array tomography is a volumetric microscopy technique that reconstructs 3D tissue structures from serial sections. This method enables detailed visualization and quantification of tissue architecture and molecular features.

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Area of Science:

Microscopy
Biotechnology
Histology

Background:

Array tomography is a volumetric microscopy method utilizing physical serial sectioning.
It involves sectioning plastic-embedded tissue, arranging sections in an array, staining, and imaging.

Purpose of the Study:

To provide a protocol for array tomography using wide-field fluorescence microscopy.
To highlight the capabilities of array tomography for visualizing and quantifying tissue structure and molecular architecture.

Main Methods:

Physical serial sectioning of plastic-embedded tissue using an ultramicrotome.
Bonding ultrathin sections into an ordered array on a coverslip.
Staining, imaging (manually or automated), and computational reconstruction into 3D volumes.

Main Results:

Enables high-quality rapid staining and imaging due to minimal section thickness.
Facilitates reliable section handling, staining, and automated imaging via the array format.
Allows repeated imaging cycles and multi-modal imaging (e.g., fluorescence and electron microscopy).

Conclusions:

Array tomography provides access to previously inaccessible features of tissue structure and molecular architecture.
Careful tissue preparation is crucial for successful array tomography.
The protocol details imaging via conventional wide-field fluorescence microscopy, with options for automation.