Della Santina Luca, Hoon Mrinalini
College of Optometry, University of Houston, Houston, TX, USA.
Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI, USA.
Methods Mol Biol. 2025;2910:179-204. doi: 10.1007/978-1-0716-4446-1_11.
Fixed tissue analyses of synaptic density and connectivity rely on analyses of image stacks containing fluorescence signals. This chapter details image analysis routines that can be implemented to determine connectivity between labeled cell types and procedures for quantification of synaptic density across a tissue volume or specifically within a labeled cell of interest. The chapter details open-source software and practices that can be implemented for these analyses as well as considerations for validation of detected signals, estimation of background noise in images, and determination of non-specific detections.
对突触密度和连接性的固定组织分析依赖于对包含荧光信号的图像堆栈的分析。本章详细介绍了可用于确定标记细胞类型之间连接性的图像分析程序,以及用于量化整个组织体积或特定感兴趣标记细胞内突触密度的程序。本章详细介绍了可用于这些分析的开源软件和实践,以及检测信号验证、图像背景噪声估计和非特异性检测确定的注意事项。