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INTRODUCTION

Optical Projection Tomography (OPT) and light sheet fluorescence microscopy (LSFM) are high resolution optical imaging techniques, ideally suited for ex vivo 3D whole mouse brain imaging. Although they exhibit high specificity for their targets, the anatomical detail provided by tissue autofluorescence remains limited.

METHODS

T1-weighted images were acquired from 19 BABB or DBE cleared brains to create an MR template using serial longitudinal registration. Afterwards, fluorescent OPT and LSFM images were coregistered/normalized to the MR template to create fusion images.

RESULTS

Volumetric calculations revealed a significant difference between BABB and DBE cleared brains, leading to develop two optimized templates, with associated tissue priors and brain atlas, for BABB (OCUM) and DBE (iOCUM). By creating fusion images, we identified virus infected brain regions, mapped dopamine transporter and translocator protein expression, and traced innervation from the eye along the optic tract to the thalamus and superior colliculus using cholera toxin B. Fusion images allowed for precise anatomical identification of fluorescent signal in the detailed anatomical context provided by MR.

DISCUSSION

The possibility to anatomically map fluorescent signals on magnetic resonance (MR) images, widely used in clinical and preclinical neuroscience, would greatly benefit applications of optical imaging of mouse brain. These specific MR templates for cleared brains enable a broad range of neuroscientific applications integrating 3D optical brain imaging.

INTRODUCTION

METHODS

RESULTS

DISCUSSION

引言

光学投影断层扫描（OPT）和光片荧光显微镜（LSFM）是高分辨率光学成像技术，非常适合用于离体全小鼠脑三维成像。尽管它们对目标具有高特异性，但组织自发荧光提供的解剖细节仍然有限。

方法

从19个经BABB或DBE透明处理的大脑获取T1加权图像，使用序列纵向配准创建一个磁共振模板。之后，将荧光OPT和LSFM图像与磁共振模板进行配准/归一化，以创建融合图像。

结果

体积计算显示经BABB和DBE透明处理的大脑之间存在显著差异，从而为BABB（OCUM）和DBE（iOCUM）开发了两个优化模板，以及相关的组织先验信息和脑图谱。通过创建融合图像，我们识别了病毒感染的脑区，绘制了多巴胺转运体和转位蛋白的表达图谱，并使用霍乱毒素B追踪了从眼睛沿视神经束到丘脑和上丘的神经支配。融合图像能够在磁共振提供的详细解剖背景下精确识别荧光信号的解剖位置。