Introduction: The confocal laser-scanning microscope (CSLM) was recently introduced. We have invented a new transmission type of double pass CSLM. This study is the first report of valuable pathological information related to bone tumor being derived using such microscopy.

Methods: The most remarkable characteristic of this microscope is the use of two laser beams twice passing through the specimen. This laser microscope can detect signals from coloring sources such hematoxylin eosin (HE) stain and obtain clear images of the organelles. The images presented here were built up as electronic signals, processed by computer analysis, and stored in frame memory. Specimens of the giant cell tumor stained with HE were examined directly by the phase contrast mode of this microscope and computer analysis was performed. Double pass CSLM and conventional microscopic views were then compared.

Results: We successfully observed sharply and sensitively positive fine granules in our laser microscopes provided higher magnification, resolution and contrast than did conventional ones. CSLM provides high magnification, contrast, resolution and can be used to observe living cells in culture in real time. With the combination of double pass CSLM and computer analysis, clear images of the subcellular organelles of various cells were successfully visualized.

Conclusion: This study suggests that double pass CSLM is an important tool for analyzing the cellular ultrastructure, physiology, and function of bone tumor. Double pass CSLM is also a powerful new instrument for orthopedics, complementing light and electron microscopy.