CAJ | 학술논문

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.
배경 조직병이학검측과정중용전통적화학고정법고정완정적안구표본역조성시망막신경섬유층적수종,영향검측결과적평개.연구표명미파고정법가이피면대측조직적손상,단조작과정중적영향인소상대연구.목적 탐색미파법고정토안시망막적괄의능량화시간삼수,평개미파고정후시망막결구적청석도화시망막적탈리정황.방법 적제22지신서란대이백토적안구,분위화학시제조、미파조화미파+화학시제조.화학시제조2지토4지안치우전통적빙작산+갑철+록방(2:1:2)혼합고정액중고정2 d;미파조8지토16지안치우400 ml수내,미파해동당분별처리80、160、240、320 s.미파+화학시제조12지토24지안미파처리후보이화학시제고정1～2 h.결과 미파조처리240 s즉가득도안조직결구염색청석적소목정-이홍염색절편,여화학시제조고정효과상사;미파+화학시제조고정적시망막결구경청석.미파조시망막탈리면적위16.3%±11.5%,화학시제조위50.0%±24.5%,미파+화학시제조위6.7%±7.8%,3조간차이균유통계학의의(F=32.433,P=0.000).미파고정200～240 s후보이화학고정1～2 h,시망막적각층결구경청석,탈리면적경소.결론 미파고정가절성시간,피면사용유해화학시제,고정효과여상규화학시제상사,병차가이방지시망막탈리;미파고정연합단잠화학고정가이득도경청석적병리절편.
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.