Background Conventional chemical method to fix the entire eyeball results in the edema of retinal nerve fiber layer and therefore influent the evaluation of tissue structure.Research showed that microwave irradiation can avoid this phenomenon. Objective This study was to investigate the appropriate energy and time duration of microwave irradiation for fixation of rabbit eyes specimen and compare the influence of different fixation methods on retinal structure. Methods Twenty-two New Zealand white rabbits were sacrificed by using venous air embolism.The eyeballs were enucleated from the rabbits and immersed in chemical reagents with glacial acetic acid,methyl aldehyde and chloroform for 2 days as the control group.The eyeballs were immersed in 400 ml physiological balance solution immediately,and then were fixed by microwave irradiation for 80,160,240 and 320 seconds under the lower power as the microwave irradiation groups.In the microwave irradiation+chemical fixation group,the specimens were immersed in chemical reagents for 1-2 hours after the microwave radiation.The morphology of retinal structure was examined and area of retinal detachment was calculated by hemotoxylin and eosin staining under the light microscope. Results A better fixation effect was obtained in the retinal section of microwave radiation for 240 seconds in comparison with that of the traditional chemical reagent fixation method for 2 days.The retina detachment ratio was 16.3%±11.5% for simple microwave method,50.0%±24.5% for chemical method,and 6.7%±7.8% for microwave+chemical method.showing a significant difference among 3 groups(F=32.43,P=0.000).Retinal staining was clear and retinal structure was almost normal in the specimens of microwave+chemical fixation group with 200-240 seeonds microwave irradiation and 1-2 hours chemical fixation. Conclusion Microwave irradiation method is a more ideal way for the fixation of retina because of taking short duration,lower toxicity and better staining.The combination of microwave radiation and chemical reagents can acquire an excellent quality of retinal section.%背景 组织病理学检测过程中用传统的化学固定法固定完整的眼球标本易造成视网膜神经纤维层的水肿,影响检测结果的评价.研究表明微波固定法可以避免待测组织的损伤,但操作过程中的影响因素尚待研究.目的 探索微波法固定兔眼视网膜的适宜能量和时间参数,评价微波固定后视网膜结构的清晰度和视网膜的脱离情况.方法 摘除22只新西兰大耳白兔的眼球,分为化学试剂组、微波组和微波+化学试剂组.化学试剂组2只兔4只眼置于传统的冰醋酸+甲醛+氯仿(2:1:2)混合固定液中固定2 d;微波组8只兔16只眼置于400 ml水内,微波解冻档分别处理80、160、240、320 s.微波+化学试剂组12只兔24只眼微波处理后辅以化学试剂固定1～2 h.结果 微波组处理240 s即可得到眼组织结构染色清晰的苏木精-伊红染色切片,与化学试剂组固定效果相似;微波+化学试剂组固定的视网膜结构更清晰.微波组视网膜脱离面积为16.3%±11.5%,化学试剂组为50.0%±24.5%,微波+化学试剂组为6.7%±7.8%,3组间差异均有统计学意义(F=32.433,P=0.000).微波固定200～240 s后辅以化学固定1～2 h,视网膜的各层结构更清晰,脱离面积更小.结论 微波固定可节省时间,避免使用有害化学试剂,固定效果与常规化学试剂相似,并且可以防止视网膜脱离;微波固定联合短暂化学固定可以得到更清晰的病理切片.
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