激光共聚焦显微镜技术在观察荧光标记和评估新骨形成中的作用

Role of confocal laser scanning microscopy in observing fluorescence labelling and evaluating new bone formation

目的旨在运用激光共聚焦显微镜光学优势，验证其活体顺序性荧光标记与透射光图像融合的特征，证实其定量骨改建的有效性。方法采用激光共聚焦显微镜与普通荧光显微镜对比观察，并用测定两种荧光标记的比率来评估新骨形成率。结果激光共聚焦显微镜技术可明显改善厚样品的图像质量；微分干涉相差与荧光图像融合，能显示荧光标记在形态结构上的精确定位，并能同时进行定性、定量和定位分布观察。结论激光共聚焦显微镜技术为评估骨代谢和骨折修复提供了一种新的手段。

Objective To verify characteristics of combination of sequential fluorescence labeling with transmitted light image by using confocal laser scanning microscopy （CLSM） and confirm its efficacy in quantitative bone formation. Methods CLSM and conventional fluorescence microscopy were employed to evaluate new bone formation by determining ratio of two fluorescence labelling. Results CLSM could significandy improve image quality of thick bone sections. CLSM was able to locate precisely the fluorescence label in morphological structure and simultaneously to determine the nature, quantity and location of the newly formed bone when differential interference contrast microscope was combined with fluorescence images. Conclusion CLSM provides a new valid method for future studies on bone mineral metabolism and evaluation of fracture repair.