Serial Block-Face SEM
Highly Automated Sectioning and Volume Data Imaging
Schematic Representation of a Typical Workflow
1
A resin-embedded sample is cut with an ultramicrotome mounted inside the SEM chamber. The exposed sample surface is imaged. This cutting and imaging process is repeated until the structure of interest is completely imaged.
2
The acquired EM images are processed and digitally aligned into a 3D data set. Cell compartments can be identified and segmented.
3
The segmented 3D data set can be visualized, investigated, and statistically analyzed.
New Discoveries from the Ultrastructure of Life Virtual Seminar Series | January – June 2024
In a series of six webinars, explore the technological underpinnings of Volume EM imaging and its growing number of application areas in neurobiology, cancer research, developmental biology, plant science, and more.
Learn about vEM-specific sample preparation and technologies (array tomography, serial block-face SEM, and FIB-SEM), advanced image processing, data analysis, and result visualization capabilities of workflow-oriented software solutions.
Application Examples
Understanding the Relationship between Structure and Function
Imaging Ultrastructural Details of Neurons
The brain is a complex organ with millions of neuronal connections and signaling pathways. Understanding the relationship between structure and function of brain tissue helps in unravelling some of this complexity to better understand how neural networks are organized and, in the long term, how to treat certain pathologies with medical interventions.
Imaging of Neurons in Cell Culture
SBF-SEM is the appropriate solution to image and follow neurons with long and thin protrusions such as dendrites and axons. Especially neurons in cell culture are difficult to image. The high proportion of the non-conductive resin makes the samples prone to charging. Focal Charge Compensation mitigates charging effects and ensures a high image quality. Ultrastructural details of neurons can be easily imaged and resolved with SBF-SEM in combination with Focal Charge Compensation.
The images show a single slice from a 3D data set of cultured hippocampal neurons expressing PSD95-APEX2 to stain post-synaptic densities (arrows). Images were acquired using a ZEISS FESEM, integrated ultramicrotome and Focal Charge Compensation. Ultrastructure such as thin dendrites and connections are visible with high resolution due to the removal of charging effects.
Single Neurons and Cellular Compartments in Mouse Brain Tissue
The video shows the cross sections of a mouse brain specimen captured using Serial Block-Face SEM. The high resolution this approach provides can be clearly seen in each of the single block-face images. Single neurons and cellular compartments can be identified and followed along the z-dimension.