Center for Molecular and Behavioral Neuroscience, Rutgers University, Newark, New Jersey, United States of America.
PLoS One. 2010 Nov 4;5(11):e13829. doi: 10.1371/journal.pone.0013829.
Two-photon scanning microscopy has advanced our understanding of neural signaling in non-mammalian species and mammals. Various developments are needed to perform two-photon scanning microscopy over prolonged periods in non-human primates performing a behavioral task. In striate cortex in two macaque monkeys, cortical neurons were transfected with a genetically encoded fluorescent calcium sensor, memTNXL, using AAV1 as a viral vector. By constructing an extremely rigid and stable apparatus holding both the two-photon scanning microscope and the monkey's head, single neurons were imaged at high magnification for prolonged periods with minimal motion artifacts for up to ten months. Structural images of single neurons were obtained at high magnification. Changes in calcium during visual stimulation were measured as the monkeys performed a fixation task. Overall, functional responses and orientation tuning curves were obtained in 18.8% of the 234 labeled and imaged neurons. This demonstrated that the two-photon scanning microscopy can be successfully obtained in behaving primates.
双光子扫描显微镜提高了我们对非哺乳动物和哺乳动物神经信号的理解。为了在执行行为任务的非人类灵长类动物中长时间进行双光子扫描显微镜，需要进行各种改进。在两只猕猴的纹状皮层中，使用 AAV1 作为病毒载体，通过基因编码的荧光钙传感器 memTNXL 转染皮质神经元。通过构建一个极其刚性和稳定的装置，该装置同时固定双光子扫描显微镜和猴子的头部，可以在长达十个月的时间内以高放大倍数对单个神经元进行长时间成像，同时最小化运动伪影。获得了高倍放大的单个神经元的结构图像。在猴子执行注视任务时，测量钙在视觉刺激过程中的变化。总的来说，在 234 个标记和成像的神经元中，有 18.8%获得了功能反应和方向调谐曲线。这表明双光子扫描显微镜可以在行为灵长类动物中成功获得。
