Observation of Bone Tissue Metabolism and Bacterial Biofilm in Aqueous Solution Using ASEM

Abstract

The ASEM ClairScope™ (JEOL Ltd.) is an inverted scanning electron microscope (SEM) (Fig. 1 left), which realizes SEM of a sample at atmospheric pressure in a readily-accessible, open container called the ASEM dish [1]. The dish has eight 0.25 mm × 0.25 mm, 100 nm-thick SiN film windows and can hold a few milliliters of culture medium for cells. It allows various types of aldehyde-fixed cells and tissues (Fig. 1 right) to be quickly observed in radical scavenger glucose solution at high resolution [1]. An optical microscope (OM) positioned above the ASEM dish can be used to observe wide areas of the sample from above, and the SEM to image specified smaller regions from below through the SiN film in the base of the dish (Fig. 1 left). The optical axes of both microscopes are aligned to image the same area in the center, and the specimen stage can be moved two-dimensionally (x-y) for targeting. The resolution measured between gold particles near the SiN film was 8nm.