[1]牟秋月,陈森耀,黄浙学,等.不同后固定时间对脑组织免疫荧光染色效果的影响[J].江西师范大学学报(自然科学版),2023,(03):242-245.[doi:10.16357/j.cnki.issn1000-5862.2023.03.04]
　MOU Qiuyue,CHEN Senyao,HUANG Zhexue,et al.The Effect of Different Postfixation Time on Immunofluorescence Staining of Brain Tissue[J].Journal of Jiangxi Normal University:Natural Science Edition,2023,(03):242-245.[doi:10.16357/j.cnki.issn1000-5862.2023.03.04]
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不同后固定时间对脑组织免疫荧光染色效果的影响()
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《江西师范大学学报》（自然科学版）[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2023年03期
页码:
242-245
栏目:
出版日期:
2023-05-25

文章信息/Info

Title:
The Effect of Different Postfixation Time on Immunofluorescence Staining of Brain Tissue
文章编号:
1000-5862(2023)03-0242-04
作者:
牟秋月; 陈森耀; 黄浙学; 潘永良*; 沈俊媛; 石依蕾; 叶梦凡
(湖州师范学院医学院,浙江省媒介生物学与病原控制重点实验室,浙江 湖州 313000)
Author(s):
MOU Qiuyue; CHEN Senyao; HUANG Zhexue; PAN Yongliang*; SHEN Junyuan; SHI Yilei; YE Mengfan
(School of Medicine,Huzhou University,Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Province,Huzhou Zhejiang 313000,China)
关键词:
快速灌注;  后固定;  催产素;  免疫荧光;  染色效果
Keywords:
rapid perfusion;  post fixation;  oxytocin;  immunofluorescence;  staining effect
分类号:
R 33
DOI:
10.16357/j.cnki.issn1000-5862.2023.03.04
文献标志码:
A
摘要:
为探讨延长后固定时间是否能改善在体快速灌注固定后脑组织的免疫荧光染色效果,该文用雄性长爪沙鼠(Meriones unguiculatus)经心脏升主动脉插入灌注针,依次快速灌注4 ℃生理盐水和质量分数为4%的多聚甲醛溶液,断头取脑,分别浸泡于质量分数为4%的多聚甲醛溶液中固定24 h和72 h后进行30 μm厚冰冻切片和免疫荧光染色,观察并比较后固定24 h和72 h下丘脑室旁核(PVN)OT免疫阳性神经元胞体数量、下丘脑前部(AH)免疫阳性神经纤维密度及其形态学结构.研究结果表明:后固定72 h组的PVN OT免疫阳性神经元胞体数量以及AH免疫阳性纤维密度均高于后固定24 h组的.与后固定24 h组相比,后固定72 h组的OT免疫阳性神经元胞体形态更加完整,境界更加清晰,神经元核周和突起免疫荧光标记强度更强.且后固定72 h组OT免疫阳性神经纤维形态比后固定24 h组的更清晰、更连续,呈串珠状.在快速灌注固定后延长后固定时间有助于改善免疫荧光染色效果.
Abstract:
To explore whether prolonging the post-fixation time can improve the immunofluorescence staining of brain tissue after rapid perfusion fixation in vivo.Male Mongolian gerbils(Meriones unguiculatus)are inserted into the perfusion needle through the ascending aorta of the heart,and then rapidly perfused with 4 ℃ normal saline and 4% paraformaldehyde solution.The brain is decapitated and postfixed in 4% paraformaldehyde solution for 24 h and 72 h respectively.Then 30 μm thick frozen sections are made and immunofluorescence staining is performed to observe and compare the number of OT immunopositive neuronal cells in the paraventricular nucleus(PVN)of the hypothalamus,the density of immunopositive nerve fibers in the anterior hypothalamus(AH)and their morphological structure after fixation for 24 h and 72 h.The number of PVN OT immunopositive neuron cell bodies and the density of AH immunopositive fibers in the 72 h postfixation group are higher than those in the 24 h postfixation group.Compared with the 24 h postfixation group, the OT immunopositive neurons in the 72 h postfixation group have more complete cell body morphology,clearer borders,and stronger immunofluorescence labeling intensity around the nucleus and neurites of neurons.Moreover,the morphology of OT immunopositive nerve fibers in the post-fixation group for 72 h is clearer,more continuous and beaded than in the post-fixation group for 24 h.Prolonged post-fixation time after rapid perfusion fixation helps to improve the effect of immunofluorescence staining.

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备注/Memo

备注/Memo:
收稿日期:2022-09-12
基金项目:国家自然科学基金(31672295)和国家级大学生创新创业训练计划(202110347050)资助项目.
通信作者:潘永良(1979—),男,浙江诸暨人,教授,博士,主要从事动物行为神经生物学研究.E-mail:02402@zjhu.edu.cn
更新日期/Last Update: 2023-05-25