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Array tomography: rodent brain fixation and embedding.

Kristina D Micheva, Nancy O'Rourke, Brad Busse

Cold Spring Harbor Protocols

|November 3, 2010

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

Area of Science:

Microscopy
Biotechnology
Cell Biology

Background:

Array tomography is a volumetric microscopy technique.
It relies on physical serial sectioning of plastic-embedded tissues.
This method allows for 3D reconstruction and analysis of tissue structures.

Purpose of the Study:

To describe the fixation and processing protocols for tissues.
To enable immunofluorescence array tomography.
To facilitate visualization of tissue structure and molecular architecture.

Main Methods:

Physical serial sectioning using an ultramicrotome.
Bonding ultrathin sections into an ordered array on a coverslip.
Staining, imaging, and computational reconstruction into 3D volumes.

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Main Results:

High-quality rapid staining and imaging are possible due to section thickness.
The array format allows for reliable handling, staining, and automated imaging.
Repeated imaging cycles and multimodal imaging (fluorescence, electron microscopy) are feasible.

Conclusions:

Array tomography provides unprecedented visualization and quantification of tissue and molecular architecture.
Careful tissue preparation is crucial for successful array tomography.
This protocol details essential steps for immunofluorescence array tomography.