Volume EM Techniques

Array Tomography

Non-destructive Volume Imaging Using Standard, Scanning Electron Microscopy

Can be performed with standard SEM
Non-destructive imaging preserves the sample
Suitable for correlative workflows

Volume EM with Array Tomography

Research groups with access to a standard SEM and some training with sample preparation can explore volume EM (vEM) with array tomography (AT). In AT, thin serial sections are cut and affixed to a sample holder. The sections are imaged by SEM and then digitally reconstructed to create a 3D data set. AT is particularly suitable for correlative workflows, such as when proteins of interest are first localized using fluorescence microscopy. A huge advantage of AT is that it does not destroy the sections, which can be archived for future use.

Schematic Representation of a Typical Workflow

1

A resin-embedded sample is cut into an array of serial sections, each with a section thickness of typically 30 – 70 nm, and attached to a sample carrier in the order they were cut.

2

Each serial section is imaged with the scanning electron microscope (SEM).

3

The acquired EM images are processed and digitally aligned into a 3D data set. Cell compartments can be identified and segmented.

4

The segmented 3D data set can be visualized, investigated, and statistically analyzed.

  
      Application Example​
      
    3D reconstruction of serial sections from root nodules with the distribution of plasmodesmata​

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Courtesy of D. Sherrier, J. Caplan, and S. Modla, University of Delaware, USA.

The Symbiotic Relationship between Plants and Bacteria

Understanding the Impact of Bacteria in Root Nodules on the Health and Condition of Plants

The root network of a plant provides access to all of the water and nutrients that are crucial components for all plant growth. Exploring the whole root network as well as understanding the influence of external microbes is important for optimizing plant health and yield. Investigating the symbiotic relationship between plants and bacteria in root nodules requires knowledge of root nodule and bacteria distribution and a combination of both fluorescence and high-resolution structural assessment is vital to understand this in detail.

Correlative array tomography enables the overlay of both fluorescence and structural data to enable visualization of root nodule and bacteria distribution.