High-frequency ultrasonic imaging of thickly sliced specimens

It has been reported that a mechanical scanning reflection acoustic microscope (hereinafter called simply "SAM"), using high frequency ultrasonic tone-burst waves, can form a horizontal cross-sectional image (i.e., c-scan image) showing a highly resolved cellular structure of biological tissue. However, the tissue prepared for the SAM has been mostly a thinly sectioned specimen. In this study, the SAM images of specimens thickly sectioned from the tissue were analyzed. Optical and scanning acoustic microscopies were used to evaluate tissues of human small intestine and esophagus. For preparing thin specimens, the tissue was embedded in paraffin, and substantially sectioned at 5similar to10mum by the microtome. For optical microscopy, the tissue was stained with hematoxylin and eosin, and affixed onto glass substrates. For scanning acoustic microscopy, two types of specimens were prepared: thinly sectioned specimens affixed on the glass substrate, wherein the specimens were deparaffinized in xylene, but not stained, and thickly sectioned specimens. Images of the thick specimens obtained with frequency at 200 MHz revealed cellular structures. The morphology was very similar to that seen in the thinly sectioned specimens with optical and scanning acoustic microscopy. In addition, scanning electron microscopy was used to compare the images of biological tissue. An acoustic lens with frequency at 200 MHz permitted the imaging of surface and/or subsurface of microstructures in the thick sections of small intestine and esophagus.