Knife-Edge Scanning Microscopy: High-Throughput Imaging and Analysis of Massive Volumes of Biological Microstructures - High-Throughput Image Reconstruction and Analysis - page 17

Figure 2.2 Tissue sectioning and imaging in KESM. (a) A close-up of the parts 2, 3, and 4 in
Figure 2.1 is shown. To the left is the microscope objective, and to the right the diamond knife
and light collimator. Submerged under water in the center is the plastic-embedded brain tissue
held in a specimen ring. (b) The principle of operation of KESM is illustrated. The objective and the
knife is held in place, while the specimen affixed on the positioning stage moves (arrow with solid
line) at the resolution of 20 nm and travel speed of 1--5, and gets scraped against the diamond
knife (5 mm wide for 10X objective), generating a thin section flowing over the knife (arrow with
solid line). Line-scan imaging is done near the very tip of the knife where the distortion is minimal
(maximum 106 m m from the tip of the knife). Illumination if provided through the diamond knife
(arrow with dashed line indicates the light path). Note that the size of the knife is exaggerated.
(Adapted from [9].)