In vivo mammalian brain Imaging using one- and two-photon fluorescence microendoscopy
In vivo mammalian brain Imaging using one- and two-photon fluorescence microendoscopy
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DOI：
10.1152/jn.00234.2004
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发表时间：
2004-11-01
影响因子：
2.5
通讯作者：
Schnitzer, MJ
中科院分区：
文献类型：
--
作者：
Jung, JC;Mehta, AD;Schnitzer, MJ
目前神经科学家可用的一套技术的主要限制之一是缺乏对活体动物大脑深处的单个细胞进行成像的方法。为了克服这一限制，我们开发了两种形式的微创荧光显微内窥镜，并测试了它们在体内成像细胞的能力。单光子和双光子荧光显微内窥镜都是基于复合梯度折射率（GRIN）透镜，其直径为350- 1，000 μ m，并提供微米级分辨率。单光子显微内窥镜允许通过眼睛或相机查看全帧图像，并且非常适合快速帧率成像。双光子显微内窥镜是一种激光扫描模式，可提供组织深处的光学切片。使用在体显微内窥镜，我们获得了视频速率电影的丘脑和CA 1海马红细胞动力学和静止帧图像的CA 1神经元和树突在麻醉大鼠和小鼠。显微内窥镜将有助于满足体内细胞成像日益增长的需求，这是由于新的合成和遗传编码荧光团的快速出现而产生的，这些荧光团可用于标记特定的大脑区域或细胞类别。
One of the major limitations in the current set of techniques available to neuroscientists is a dearth of methods for imaging individual cells deep within the brains of live animals. To overcome this limitation, we developed two forms of minimally invasive fluorescence microendoscopy and tested their abilities to image cells in vivo. Both one- and two-photon fluorescence microendoscopy are based on compound gradient refractive index (GRIN) lenses that are 350-1,000 mum in diameter and provide micron-scale resolution. One-photon microendoscopy allows full-frame images to be viewed by eye or with a camera, and is well suited to fast frame-rate imaging. Two-photon microendoscopy is a laser-scanning modality that provides optical sectioning deep within tissue. Using in vivo microendoscopy we acquired video-rate movies of thalamic and CA1 hippocampal red blood cell dynamics and still-frame images of CA1 neurons and dendrites in anesthetized rats and mice. Microendoscopy will help meet the growing demand for in vivo cellular imaging created by the rapid emergence of new synthetic and genetically encoded fluorophores that can be used to label specific brain areas or cell classes.