解剖学报

解剖学报 ›› 2015, Vol. 46 ›› Issue (1): 118-121.doi: 10.16098/j.issn.0529-1356.2015.01.021

• 技术方法 • 上一篇    下一篇

用于膜片钳记录的脑片制备方法

陈醒1 刘晓栋2 李琳3 王珂4 白文佩1* 秦丽华4 于龙川2*   

  1. 1.北京大学第一医院妇产科,北京 100034; 2.北京大学生命科学学院神经生物学实验室，生物膜与膜生物工程国家重点实验室,北京 100871; 3.北京大学第一医院实验中心，北京 100034; 4.北京大学医学部解剖学与组织胚胎学系，北京 100191
  • 收稿日期:2014-07-01 修回日期:2014-08-10 出版日期:2015-02-06 发布日期:2015-02-06
  • 通讯作者: 白文佩,于龙川 E-mail:bwp66@163.com
  • 基金资助:

    低雌激素经5-羟色胺通路致体温调节中枢功能紊乱的机制

Exploring the method of preparing brain slices for patch clamp recording

CHEN Xing1 LIU Xiao-dong2 LI Lin3 WANG Ke4 BAI Wen-pei 1* QIN Li-hua4 YU Long-chuan 2*   

  1. 1.Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing 100034,China; 2. Laboratory of Neurobiology and State Key Laboratory of Biomembrane and Membrane Biotechnology, College of Life Sciences,Peking University,Beijing 100871,China 3. Department of Central Laboratory, 
    Peking University First Hospital, Beijing 100034,China 4. Department of Anatomy and Embryology, Peking University Health Science Center, Beijing 100191,China
  • Received:2014-07-01 Revised:2014-08-10 Online:2015-02-06 Published:2015-02-06
  • Contact: Bai Wen-pei ,YU Long-chuan E-mail:bwp66@163.com

摘要：

目的 探索用于膜片钳记录的脑片制备方法并分析鼠龄、溶液、操作过程等因素对神经细胞活性的影响。 方法 取各年龄段的Sprague-Dawley大鼠60只，麻醉后快速断头取脑，冰镇后取出并修块，用振动切片机切取300μm下丘脑冠状脑片，人工脑脊液33℃孵育45min后，在红外微分干涉相差显微镜下观察脑片神经元。
结果 镜下可观察到状态良好的神经元（边界清晰、立体感强、细胞核和颗粒不可见），神经元活性的好坏与实验条件和实验操作有关。 结论 鼠龄、溶液、实验操作等因素与脑片神经元活性密切相关，实验过程中需要严格控制各种实验条件。

关键词: 脑片, 神经元, 膜片钳, 大鼠

Abstract:

  Objective To explore a method of preparing brain slices for patch clamp technique and to analyze effects of age,solutions and processes of operation on neurons of brain slices. Methods Sixty Sprague-Dawley rats were anesthetized and quickly decapitated. Their brains were cooled and dissected then 300μm coronary brain slices were cut and incubated in the artifical cerebrospinal fluid at 33℃ for 45 minutes. Neurons were observed under an infrared differential aberration interference microscope. Results Neurons showed clear boundary, and three-dimensional appearance and nuclei and particles were not seen. The activity of brain slices depended on various experiment conditions.Conclusion The activity of neurons in the brain slice is associated with age,solutions and processes of operation, thus in experiments we should strictly control these factors.

Key words: Brain slice, Neuron, Patch clamp, Rat