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.

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

• JEOL JSM 7800F LV with Gatan 3View2XP.