3D Histology Using the Synchrotron Radiation Propagation Phase Contrast Cryo-microCT / 체질인류학회지

3D histology is a imaging system for the 3D structural information of cells or tissues. The synchrotron radiation propagation phase contrast micro-CT has been used in 3D imaging methods. However, the simple phase contrast micro-CT did not give sufficient micro-structural information when the specimen contains soft elements, as is the case with many biomedical tissue samples. The purpose of this study is to develop a new technique to enhance the phase contrast effect for soft tissue imaging. Experiments were performed at the imaging beam lines of Pohang Accelerator Laboratory (PAL). The biomedical tissue samples under frozen state was mounted on a computer-controlled precision stage and rotated in 0.18° increments through 180°. An X-ray shadow of a specimen was converted into a visual image on the surface of a CdWO4 scintillator that was magnified using a microscopic objective lens (X5 or X20) before being captured with a digital CCD camera. 3-dimensional volume images of the specimen were obtained by applying a filtered back-projection algorithm to the projection images using a software package OCTOPUS. Surface reconstruction and volume segmentation and rendering were performed were performed using Amira software. In this study, We found that synchrotron phase contrast imaging of frozen tissue samples has higher contrast power for soft tissue than that of non-frozen samples. In conclusion, synchrotron radiation propagation phase contrast cryo-microCT imaging offers a promising tool for non-destructive high resolution 3D histology.

The effect of nerve preservation methods on rat sciatic nerve structures studied with Synchrotron small-angle X-ray scattering (SAXS)

BACKGROUND: Synchrotron small-angle X-ray scattering (SAXS) is a very useful technique for experimental study of the nano-structure of the nervous system of animals. The study was designed to evaluate nerve preservation methods for the measurement of SAXS patterns. METHODS: Normal sciatic nerves extracted from male Sprague- Dawley rats were preserved in saline (N = 2), formalin (N = 2) or liquid nitrogen (N = 2) for 1 day, followed by measurement of SAXS patterns. SAXS patterns of normal sciatic nerves (N = 3) extracted just before the initiation of the experiment were used as controls. The study was carried out using the 4C1 beamline at Pohang Accelerator Laboratory in Korea. Incoming X-rays were monochromatized at 11 keV using a double multilayer (WB4C) monochromator with beam size of approximately 0.5 (V) × 0.8 (H) mm2. The exposure time was set at 60 sec, and 8 to 12 images per sample were acquired at a 0.5 mm interval. RESULTS: The periodic peaks of interfibrillar space between collagen fibrils were undetectable. The periodic peaks of the myelin sheath and collagen fibers were weakly detected or undetected in the nerves preserved in normal saline or formalin. The periodic peaks and intensity of the myelin sheath, collagen fibers, and interfibrillar space between collagens in the nerves preserved in liquid nitrogen were comparable to those of nerves in the ex vivo state. CONCLUSIONS: The study results indicated that preservation of nerves in liquid nitrogen is adequate for measurements with SAXS. However, saline and formalin preservation techniques were inadequate for SAXS measurement.