光波-微波辐射技术在骨组织快速脱钙与免疫组化染色中的应用

Application of Light Wave and Microwave Radiation Techniques in Rapid Decalcification and Immunohistochemistry of Osseous Tissues

目的探讨光波-微波辐射技术在骨组织快速脱钙与免疫组化染色中的作用。方法用8%硝酸分别在单纯微波、单纯光波、光波-微波组合Ⅰ和光波-微波组合Ⅱ的条件下脱钙,同时以室温脱钙作为对照;应用光波-微波-LSAB免疫组织化学染色技术检测不同脱钙条件下ANP、CD31、CD34、5-HT、S-100、NF、GFAP和Vimentin的表达。结果8%硝酸室温脱钙时间最长,光波-微波组合脱钙时间最短;微波组、光波组、光波-微波组合Ⅰ组、Ⅱ组脱钙的免疫组化染色均明显比对照组好;光波-微波组合Ⅰ组、Ⅱ组脱钙的免疫组化染色均明显优于单纯微波、单纯光波法;光波-微波组合Ⅱ-LSAB免疫组织化学染色技术比常规免疫组织化学染色所用时间短、染色效果好。结论光波-微波辐射脱钙及免疫组化染色所用的时间明显缩短,染色效果好。

Objective To investigate the effect of light wave-microwave radiation techniques in decalcification and immunohisochemistry of osseous tissues. Methods Decalcification with 8% nitric acid （NA） was carried out under conditions of microwave group, light wave group, light wave-microwave combination （LMC） group Ⅰ and LMC group Ⅱ , respectively, and decalcification with 8% nitric acid carried out at room temperature （20℃） was used as control group. The expression of ANP, CD31 ,CD34,5-HT, S-100, NF, GFAP and vimentin was detected by light wave-microwave LSAB method under different decalcification conditions. Result The nitric acid decalcification time at room temperature was the longest, up to 36 - 48 hours, and that of light wave-microwave Ⅰ and Ⅱ methods was the shortest, 40 - 50 rain and 35 - 45 min, respectively. The effect of immtmohistochemical staining with microwave, light wave, LMC Ⅰ and Ⅱ methods was better than that obtained in the control group. The results of immunohistochemical staining after LMC Ⅰ and Ⅱ method were apparently better than that obtained by simple microwave or light wave treatment. Immunohistochemial staining after LMC Ⅱ treatment needed shorter staining time and showed better staining result than conventional immunohistochemistry. Conelusion The application of decalcification combined with light wave-microwave radiation technique can significantly shorten the immunohistochemical staining process of osseous tissues and demonstrate better staining results.