微波固定组织对超微结构影响的电镜观察

Electron microscopic observation on effect of ultrastructure of fixed tissue with microwave

目的 :观察微波照射 (microwaveirradiation ,MI)固定对组织细胞器结构的影响。方法 :将大鼠肝组织分别浸泡在生理盐水、0 2 5 %戊二醛、2 5 %戊二醛、5 %戊二醛液体中 ,按不同时间进行微波照射固定后 ,常规制片 ,电镜观察。结果 :MI时间≤2 0s,液体的温度≤ 2 5℃ ,肝细胞内各有形成分均有一定膨胀。MI时间≥ 6 0s,液体的温度≥ 45℃时 ,肝细胞内各有形成分示不同程度破坏 ,其中戊二醛各组均较生理盐水组严重 ,且戊二醛组中 ,戊二醛浓度越高 ,对细胞内结构破坏性就越大。微波照射在 40s,各组液体的温度为 33℃左右时固定最好 ,超微结构清晰 ,细胞器中各有形成分膨胀及损伤最小。结论 :用微波照射浸泡在生理盐水内固定组织 ,做常规电镜制片 ,不失为一种组织固定的较佳方案。

Purpose To observe the effect of microwave irradiation on cell organelles in tissue. Methods Four groups of mouse hepatic tissue immersed in physiological saline, 0 25%、2 5%、5% glutaraldehyde were irradiated with microwave for various time, then it was slided using normal method to observe by electron microscope. Results The hepatic cell organelles such as mitochondrion, endoplasmic reticulum were swelled to vavious degree when the irradiating time was less than 20 seconds and the solution temperature was lower than 25℃. When the irradiation time was more than 60 seconds and the solution temperature was higher than 45℃, the hepatic cell organelles were damaged to various degree. The destructive effect on hepatic cell organelles in glutaraldehyde group was higher than in physiological saline group.In glutaraldehyde groups ,destructive effect was remarkably gerious with its concentration. When the irradiation time was 40 seconds and the solution temperature was 33℃ or so ,the best fixed effect could be acquired. The ultrastructure was clear and no swelling or damage could be seen in cell organelles. Conclusion It is a better method that tissue fixation using physiological saline with micowave irradiation instead of traditional glutaraldehyde solution.