Microtome SEM

Serial Block Face Imaging, SBFI, using in-situ microtome. This easy to install and remove 3View unit allows for in-situ SEM microtome cut slices from 200nm to 15nm thickness.

The cutting is done by means of the ultra-microtome positioned in the SEM chamber[2]. The process is followed by creating a three-dimensional orientation dataset of the selected volume. The SBFI method allows easy collection of huge amount of quantitative data and overcomes the bottleneck of traditional composition of transmission electron microscopy data with extremely demanding sample preparation and demanding data mining in general. The SBFI technique provides the unique insight into the cell and/or tissue architecture and enables yielding of the crucial knowledge for understanding of pato/ physiological functions on all levels of an organism.

Volume reconstruction of a mouse liver tissue as obtained from 500 slices, 20 nm thick, and field of view 65 μm, HiVac. The green spherical caps show the cells nuclei, the red protrusions represent sinusoids. The dataset is processed in ORS software. Sample was provided by Gatan Inc.

The human embryonic stem cell colony acquired in BSE mode during SBFI. UniVac mode; 50 Pa, 5 kV, field of view 65 μm. Sample by courtesy of Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno

Both images above at LowVac 30 Pa using Back Scatter detector at 2.5 keV beam energy. Field of View:10um.

Both images above at LowVac 30 Pa using Back Scatter detector at 2.5 keV beam energy.