中山大学学报自然科学版 ›› 2011, Vol. 50 ›› Issue (5): 110-113.

• 研究论文 • 上一篇    下一篇

活体小鼠脑组织Ca2+敏感荧光染料双光子成像的观察

彭诗宇1,任振华2,徐金勇2,顾怀宇3,李光武1,2   

  1. (1.安徽医科大学神经生物学研究所,安徽 合肥230032;2. 安徽医科大学人体解剖学教研室,安徽 合肥 230032;3.中山大学中山医学院人体解剖学教研室,广东 广州 510080)
  • 收稿日期:2011-01-05 修回日期:1900-01-01 出版日期:2011-09-25 发布日期:2011-09-25
  • 通讯作者: 李光武

Research on Ca2+ Sensitive Fluorescent Indicators for Two-Photon Brain Imaging in Live Mouse

PENG Shiyu1, XU Jinyong2, REN Zhenhua2, GU Huaiyu3, LI Guangwu1,2   

  1. (1. Institution of Neurobiology, Anhui Medical University, Hefei 230032,Chian; 2. Department of anthropotomy, Anhui Medical Unoversity, Hefei 210032,China;3. Department of anthropotomy, Sun Yatsen University,Guangzhou 510080,China)
  • Received:2011-01-05 Revised:1900-01-01 Online:2011-09-25 Published:2011-09-25

摘要: 使用双光子激光共聚焦显微镜对活体小鼠脑组织(体感皮层)进行形态学和细胞内Ca2+变化水平的观察和记录。方法是对小鼠开颅后向脑内注射Ca2+敏感荧光染料Oregon Green 488 BAPTA-1乙酰氧基甲酯,然后在双光子显微镜下观察并记录。双光子显微镜下的图像清晰生动,可扫描到大脑皮层以下较深区域,并能检测神经元细胞内Ca2+离子即时水平变化。实验结果表明通过注射Ca2+敏感荧光染料并结合多光子激光共聚焦显微镜观察活体动物脑部变化的方法可以得到高分辨率图像并且作为分析的来源。

关键词: 活体脑成像, 双光子激光共聚焦显微镜, Ca2+敏感染料, 体感皮层

Abstract: To observe the brain image and the change of Ca2+ level in live mouse by using twophoton laser scanning confocal microscope. Using craniotomy to expose target area of mouse brain(somatosensory cortex) then injectCa2+ sensitive fluorescent indicator,for instance, Oregon Green 488 BAPTA-1 acetoxymethyl. After that, depose the mouse under the twophoton to observe and record. The brain image record by twophoton is clear and vivid .And photograph indicate that by using twophoton confocal can reach deeper area of the brain and record the fast change of Ca2+ in single neuron. The results show that by using multiphoton confocal microscope couple with calcium sensitive fluorescent indicator can get high resolution image of brain of live mouse.

Key words: brain image in vivo, Two-photon laser scanning confocal microscope, Ca2+ sensitive dye, somatosensory cortex

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