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The confocal laser scanning microscope (CLSM) has become the most recently developed microscopic techniques in biological research. We have tried to assess the utility of the confocal imaging in the morphological observation of the hard tissues. The more improvement in resolution was demonstrated by the CLSM image of the incorporation of the rhodamine-labeled albumin into the rabbit incisor dentin compared with the normal fluorescence image. It provided the observation at high resolution and quality of structures deep within the trabecular bone of the rat tibia labeled with calcein and gave serial optical sections by changing the plane of focus through the thick specimen. A fully three-demensional representation of the bone was easily obtained by the computer-generated analysis of the stored images. The cytoskeletal components of the isolated rat osteoclasts were clearly visible in the immuno-fluorescence CLSM images. The high performance images of the CLSM are valuable for studying the morphology of the hard tissues. Moreover the application of the CLSM should lead to significant advances in our understanding of the function of the hard tissues.

Morphological observations of the hard tissues with the confocal laser scanning microscope.