Extended field-of-view ultrathin microendoscopes for high-resolution two-photon imaging with minimal invasiveness.
Extended field-of-view ultrathin microendoscopes for high-resolution two-photon imaging with minimal invasiveness.
复制标题
扩展的视野超明显微型镜检查，用于高分辨率两光子成像，具有最小的侵入性。
DOI：
10.7554/elife.58882
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发表时间：
2020-10-13
期刊：
影响因子：
7.7
通讯作者：
Fellin T
中科院分区：
文献类型：
--
作者：
Antonini A;Sattin A;Moroni M;Bovetti S;Moretti C;Succol F;Forli A;Vecchia D;Rajamanickam VP;Bertoncini A;Panzeri S;Liberale C;Fellin T
在整个视野（FOV）内以高且均匀的空间分辨率成像神经元活动，并且在脑深部区域中具有有限的侵入性，这是神经科学进步的基础，但也是一个重大的技术挑战。我们通过使用通过3D微打印生成的非球面微透镜校正基于梯度折射率透镜的微内窥镜（≤500 µm）中的光学像差来实现这一目标。校正后的显微内窥镜具有扩展的FOV（eFOV），具有用于双光子荧光成像的均匀空间分辨率，并且不需要修改光学设置。合成钙成像数据显示，与未校正的内窥镜相比，eFOV显微内窥镜改善了信噪比，并更精确地评估了相关神经元活动。我们在清醒的头部固定小鼠中实验验证了这些预测。此外，使用eFOV显微内窥镜，我们证明了细胞特异性编码的行为状态依赖的信息在分布的功能子网络中的初级躯体感觉丘脑核。因此，eFOV显微内窥镜是用于深度双光子功能成像的小横截面即用型工具，具有前所未有的高和均匀的空间分辨率。
Imaging neuronal activity with high and homogeneous spatial resolution across the field-of-view (FOV) and limited invasiveness in deep brain regions is fundamental for the progress of neuroscience, yet is a major technical challenge. We achieved this goal by correcting optical aberrations in gradient index lens-based ultrathin (≤500 µm) microendoscopes using aspheric microlenses generated through 3D-microprinting. Corrected microendoscopes had extended FOV (eFOV) with homogeneous spatial resolution for two-photon fluorescence imaging and required no modification of the optical set-up. Synthetic calcium imaging data showed that, compared to uncorrected endoscopes, eFOV-microendoscopes led to improved signal-to-noise ratio and more precise evaluation of correlated neuronal activity. We experimentally validated these predictions in awake head-fixed mice. Moreover, using eFOV-microendoscopes we demonstrated cell-specific encoding of behavioral state-dependent information in distributed functional subnetworks in a primary somatosensory thalamic nucleus. eFOV-microendoscopes are, therefore, small-cross-section ready-to-use tools for deep two-photon functional imaging with unprecedentedly high and homogeneous spatial resolution.