Application of a Newly Developed Real-Time 3D Scanning Electron Microscope to the Biomedical Research
Abstract: The stereoscopic visualization has been a matter of interest for researchers using the scanning electron microscope (SEM). For this purpose, two images of the same area with a small tilt angle difference are usually collected by sample tilting. However, we recently developed an novel SEM, which can obtain real-time stereo-images by using a beam-tilting method. In our real-time 3D SEM, each line is recorded twice with rocking beam angles during scanning, resulting in the simultaneous collection of two images with different tilt angles. To obtain high-resolution images, the off-axis aberration formed by off-axis beam deflection is corrected by inserting an additional lens. The obtained stereo images are displayed on a 3D liquid crystal display, which is also designed for representation of stereo images without using any special glasses. Thus, we could obtain stereo SEM images of various biological tissue samples in real-time at different magnifications even by naked eyes. We then installed compact manipulators in our SEM, and dissected tissue blocks during SEM operation. Because the spatial distance between the sample and needle could be recognized accurately and stereoscopically in our SEM, we could dissect samples more easily than before.
Key words: Scanning electron microscopy, stereo-imaging, 3D liquid crystal display, biological samples, manipulation