Applications for Array Tomography

The brain is a complex organ with millions of neural connections and signalling pathways. Understanding the relationship between structure and function of neural tissue helps in unravelling some of this complexity to better understand how the brain works and in the long term how to treat malfunctions with medical interventions.

ZEISS Atlas 5 Array Tomography is a hardware/software tool that can automatically image the serial sections on the substrate down to nanometer resolution. This unique workflow is easy-to-use and has been specifically designed for automated imaging to enable the 3D visualization of the large volumes necessary for understanding the vast connections in brain tissue.

This image is a large-scale SEM image of an ultrathin section of a mouse brain mounted on a substrate. It has been acquired using Atlas 5 Array Tomography to assist in automating the acquisition.

Subcellular details such as the nucleus and mitochondria can be identified and when reconstructed in 3D the connections between the different neurons can be mapped.

This video demonstrates the great value of using Atlas 5 Array Tomography to streamline your AT workflows. As the video progresses you can see how high-resolution data is captured from each slice can be combined to ultimately generate a 3D dataset.

When imaging brain using Array Tomography you have many hundreds and sometimes thousands of slices available that need to be located and imaged. Manual identification of each of these slices is laborious and time consuming. Atlas 5 automatically and quickly identifies each of the slices such that subsequent imaging can begin quickly and efficiently and all of the high-resolution information about each slice is registered in the software.

The root network of a plant provides access to 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.

The images are either 3D reconstructions or single slices of fluorescence data from serial sections from root nodules showing the distribution of plasmodesmata. The stack can be overlaid with the SEM data to investigate the relationship of bacterial infection and distribution of plasmodesmata.

The sample was embedded in Epon and cut with an ultramicrotome. The serial sections were transferred on ITO-coated cover glasses by means of a micromanipulator. Cell walls were stained with Calcofluor White (blue) and the plasmodesmata were stained against Callose with Alexa Fluor 647. The sample was post-stained for imaging in the SEM.

Huntington’s Disease is an incurable, progressive neurodegenerative disease that is caused by a defective gene on chromosome 4 that codes for the huntingtin protein. The defective gene leads to the formation of mutant huntingtin protein which misfolds and aggregates causing protein plaque development in the brain. Such deterioration in the brain leads to disruption of movement, behavior, thinking and emotions.
  

The correlation of high-resolution ultrastructural information and fluorescence data is important to thoroughly understand the biology and progression of Huntington’s disease.

The image on the left/above show a schematic representation of the information gained in 3D through combining the serial LM and SEM images of macrophages expressing the protein huntingtin resulting from a ZEISS ZEN Correlative Array Tomography experiment.