Specimen Preparation, Imaging, and Analysis Protocols for Knife-edge Scanning Microscopy

A simple but precise method for quantitative measurement of the quality of the laser focus in a scanning optical microscope

We report a method for characterizing the focussing laser beam exiting the objective in a laser scanning microscope. This method provides the size of the optical focus, the divergence of the beam, the ellipticity and the astigmatism. We use a microscopic-scale knife edge in the form of a simple transmission electron microscopy grid attached to a glass microscope slide, and a light-collecting op...

Knife-Edge Scanning Microscopy: High-throughput Imaging and Analysis of Massive Volumes of Biological Microstructures

Multiscale Exploration of Mouse Brain Microstructures Using the Knife-Edge Scanning Microscope Brain Atlas

Connectomics is the study of the full connection matrix of the brain. Recent advances in high-throughput, high-resolution 3D microscopy methods have enabled the imaging of whole small animal brains at a sub-micrometer resolution, potentially opening the road to full-blown connectomics research. One of the first such instruments to achieve whole-brain-scale imaging at sub-micrometer resolution i...

Specimen Preparation for Electron Microscopy

In general the SEM sample preparation techniques are more or less similar to the metallographic sample preparation technique for optical microscopy, because both the microscopes reveals the surface topography of sample. In SEM a focussed beam of electrons scans over the specimen surface and the electrons emitted from the sample surface controls the brightness of the CRT spot which also scans th...

Fast macro-scale transmission imaging of microvascular networks using KESM

Accurate microvascular morphometric information has significant implications in several fields, including the quantification of angiogenesis in cancer research, understanding the immune response for neural prosthetics, and predicting the nature of blood flow as it relates to stroke. We report imaging of the whole mouse brain microvascular system at resolutions sufficient to perform accurate mor...