为探讨原位条件下关节软骨及下骨组织成分含量变化等深层次微观信息， 采用拉曼光谱技术结合组织光透明技术来研究软骨组织的光透明效果。 选用甘油、 碘海醇作为光透明剂， 对在不同光透明剂浓度和浸渍时间下的犬膝关节股骨端软骨样本进行拉曼光谱采集。 通过计算磷酸基团(920~960 cm-1 ) 和酰胺Ⅰ带(1 595~1 700 cm-1)的积分强度， 获得拉曼强度比（磷酸基团/酰胺Ⅰ）， 探索在不同浸渍时间（10~60 min）、 不同浓度下（甘油40%， 60%， 80%和100%， 碘海醇50， 150， 250和350 mg·mL-1）组织光透明的规律， 即寻找合适的透明浓度和透明时间。 结果显示： 相比于无透明剂情形， 两种透明剂均使该强度比信号增强。 在同一时间范围内， 甘油和碘海醇分别在60%和150 mg·mL-1 浓度下能获得较好的透明效果； 而在不同浓度下， 甘油的透明效果均在20 min最强， 而碘海醇的透明效果一般是在50 min后开始增强。 在光透明剂的作用下激发光透过软骨可以直接探测到软骨下骨的拉曼光谱信息， 这为研究骨关节炎深层次病因提供了方法和思路。

To investigate deeper microscopic information of articular cartilage and subchondral bone components in situ, Raman spectroscopy combined with tissue optical clearing technique was used to study the clearing effect of articular cartilage. Raman spectra of the cartilage adhered on femur ends in canine knees were collected after they were impregnated at different time and different concentrations of clearing agents, glycerol and iohexol. The integrated Raman intensity ratio of phosphate group (920~960 cm-1) to the amide Ⅰ band (1 595~1 700 cm-1) was obtained to seek the rule of tissue optical clearing at different impregnating time (10～60 min) and different concentrations of clearing agents (40%, 60%, 80% and 100% for glycerol, 50, 150, 250, 350 mg·mL-1 for iohexol), which is to get the optimum concentration and time. It was found that the both agents could enhance the Raman signal compared to the case of no clearing agent. With equal time for impregnating, glycerol and iohexol were able to obtain good results at 60% and 150 mg·mL-1, respectively. Under different concentrations, the clearing effect of glycerol was the strongest in 20 min, while that of iohexol was enhanced after 50-min impregnation in general. In conclusion, the Raman scattering information of subchondral bone can be directly detected when the clearing agents penetrating through cartilage, which will provide a novel way and a potential solution into study on osteoarthritis etiology in depth.