| 摘要: |
| 目的 探讨制作高质量大鼠脑组织连续石蜡切片的方法。 方法 大鼠麻醉后经灌注固定取脑组织，再定向3mm切块浸泡固定；梯度酒精脱水，其中95%乙醇2次各1h，无水乙醇2次各30min；二甲苯透明2次各10min；浸蜡先后采用熔点56~58℃石蜡60℃1h、58~60℃石蜡60℃2h；包埋石蜡与第二道浸蜡相同。新刀4μm切片，42℃摊片，防脱载玻片贴附切片。 结果 脑组织灌注固定后呈乳白色，有一定韧性；二甲苯透明后呈透明状，不混浊；切片厚度计数约80μm后，可获得高质量连续切片。HE染色切片质量良好，镜下组织结构完整，核质清楚；免疫组织化学染色较少脱片。 结论 灌注固定、脱水透明时间、浸蜡包埋选择、摊片温度及防脱玻片等，是制备高质量大鼠脑组织连续石蜡切片的关键。 |
| 关键词: 脑 大鼠 石蜡切片 高质量 方法 |
| DOI：10.16781/j.0258-879x.2016.04.0527 |
| 投稿时间：2015-06-23修订日期：2015-09-14 |
| 基金项目: |
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| Preparation of high-quality successive paraffin sections of rat brain tissue |
| CHU Ying-xin1,ZHANG Shu-bo1,GAO Xiao-zeng1,CHENG Guang2,ZHANG Xiao-ping1* |
(1. Department of Anesthesiology, Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, Hebei, China;
2. The Central Lab, Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, Hebei, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the methods of preparing high-quality successive paraffin sections of rat brain tissue. Methods Rats were anesthetized and transcardiac perfusion fixation was performed for collecting brain tissue. Then the brain was sequentially cut into 3mm-thick blocks and immersed in fixative; dehydrated in a gradient ethanol series, with 95% ethanol 2 times for 1 hour each, and ethanol 2 times for 30 minutes each; cleared with xylene 2 times for 10 minutes each; dipped wax at 60℃ with paraffin of melt point 56~58℃ for 1 hour then followed by melt point 58~60℃ for 2 hours; and embedded with the same paraffin as the second waxdip. 4μm sections were sliced with new knives, flattened with 42℃ water bath, and attached with adhesion slides. Results After perfusion fixation the brain tissue appeared milk-white and had certain toughness; through xylene clearing the brain presented totally transparent, without any cloudy structure; and at thickness counting approximate 80μm, we could harvest high-quality consecutive sections. The result of HE staining turned favourable, the microscopic histological structure were intact, cell nucleus and plasma were fresh; and there was also less section tissue fading during immunohistochemistry staining. Conclusion Perfusion fixation, time of dehydration and clearing, selection of waxdip and embedding, temperature of flattening water bath and the use of adhesion slides, are the key factors to the preparation of high-quality successive paraffin sections of rat brain tissue. |
| Key words: brain rats paraffin sections high quality methods |
