Gubra ApS, 2970, Hørsholm, Denmark.
Department of Applied Mathematics and Computer Science, Technical University Denmark, 2800, Kongens Lyngby, Denmark.
Neuroinformatics. 2021 Jul;19(3):433-446. doi: 10.1007/s12021-020-09490-8.
In recent years, the combination of whole-brain immunolabelling, light sheet fluorescence microscopy (LSFM) and subsequent registration of data with a common reference atlas, has enabled 3D visualization and quantification of fluorescent markers or tracers in the adult mouse brain. Today, the common coordinate framework version 3 developed by the Allen's Institute of Brain Science (AIBS CCFv3), is widely used as the standard brain atlas for registration of LSFM data. However, the AIBS CCFv3 is based on histological processing and imaging modalities different from those used for LSFM imaging and consequently, the data differ in both tissue contrast and morphology. To improve the accuracy and speed by which LSFM-imaged whole-brain data can be registered and quantified, we have created an optimized digital mouse brain atlas based on immunolabelled and solvent-cleared brains. Compared to the AIBS CCFv3 atlas, our atlas resulted in faster and more accurate mapping of neuronal activity as measured by c-Fos expression, especially in the hindbrain. We further demonstrated utility of the LSFM atlas by comparing whole-brain quantitative changes in c-Fos expression following acute administration of semaglutide in lean and diet-induced obese mice. In combination with an improved algorithm for c-Fos detection, the LSFM atlas enables unbiased and computationally efficient characterization of drug effects on whole-brain neuronal activity patterns. In conclusion, we established an optimized reference atlas for more precise mapping of fluorescent markers, including c-Fos, in mouse brains processed for LSFM.
近年来，全脑免疫标记、光片荧光显微镜（LSFM）以及随后的数据与通用参考图谱的配准相结合，使得对成年小鼠大脑中的荧光标记物或示踪剂进行 3D 可视化和定量成为可能。如今，Allen 脑科学研究所（Allen Institute of Brain Science，AIBS）开发的共同坐标框架版本 3（Allen Institute of Brain Science，AIBS CCFv3）被广泛用作 LSFM 数据配准的标准脑图谱。然而，AIBS CCFv3 是基于与 LSFM 成像不同的组织处理和成像方式构建的，因此，数据在组织对比度和形态上存在差异。为了提高 LSFM 成像全脑数据的配准和定量的准确性和速度，我们基于免疫标记和溶剂清除的大脑创建了一个优化的数字小鼠脑图谱。与 AIBS CCFv3 图谱相比，我们的图谱在使用 c-Fos 表达测量的神经元活性的映射方面更快、更准确，尤其是在后脑。我们通过比较在瘦鼠和饮食诱导肥胖鼠中急性给予 semaglutide 后 c-Fos 表达的全脑定量变化，进一步证明了 LSFM 图谱的实用性。与 c-Fos 检测的改进算法相结合，LSFM 图谱能够对药物对全脑神经元活动模式的影响进行无偏和计算高效的特征描述。总之，我们为 LSFM 处理的小鼠大脑中的荧光标记物（包括 c-Fos）的更精确映射建立了一个优化的参考图谱。
