Serial block face scanning electron microscopy (SBFSEM) can generate large 3D volumes by assembling sequential 2D images acquired in a scanning electron microscope.

SBFSEM incorporates a microtome within the scanning electron microscope chamber. The microtome cuts a thin section from the resin specimen block, an image is collected on the backscatter electron detector, and the procedure is repeated until the required number of images to reconstruct a 3D volume of the sample is generated.

This technique uses room temperature processing techniques with high levels of en bloc heavy metal staining required to generate a sufficiently robust back-scattered electron signal to form an image.

Micrographs of individual synapses
Micrographs of individual synapses
Examples of use

We have used this technique to:

  • investigate changes in cell organisation and structure in a developing zebrafish
  • understand the association of intracellular and synaptic organisation in cochlear inner hair cells and changes attributable to noise or age-induced hearing loss
  • link ultrastructural organisation to distance-dependent calcium signals along tapering dendrites
  • investigate 3D organisation and volume changes in mitochondria in normal and diseased cardiac tissue
Equipment available
Images
Micrographs of individual synapses
Micrographs of individual synapses

Micrographs of individual synapses with the postsynaptic compartment in yellow and presynaptic in green.