Sholl analysis of mitochondria from article The adaptor protein Miro1 modulates horizontal transfer of mitochondria in cancer

The following script was used to quantify mitochondrial distribution (Sholl analysis macro) using FIJI (ImageJ) software in the context of the knockout of the Miro1 mitochondrial adaptor protein as described previously in López‐Doménech et al. with minor modifications [1]. A brief description of the script's function follows (the script, demonstration data, and expected output files are attached). On the input files (z-stack confocal images of a single cell containing three channels corresponding to the nucleus, actin network, and mitochondria), the nucleus is segmented and delineated to retrieve the first shell. Subsequently, the area of the segmented actin network or segmented mitochondria is quantified in 30 concentric shells, each increased by one micrometre in radius. Output data are saved as a .csv file plus an overview image to double-check successful segmentation. [1] López-Doménech G, Covill-Cooke C, Ivankovic D, Halff EF, Sheehan DF, Norkett R, Birsa N, Kittler JT. Miro proteins coordinate microtubule- and actin-dependent mitochondrial transport and distribution. EMBO J. 2018 Feb 1;37(3):321-336. doi: 10.15252/embj.201696380. Epub 2018 Jan 8. PMID: 29311115; PMCID: PMC5793800.本脚本用于在敲除Miro1线粒体衔接蛋白（Miro1 mitochondrial adaptor protein）的实验背景下，借助FIJI（ImageJ）软件量化线粒体分布（采用Sholl分析宏程序（Sholl analysis macro）），相关实验方法参照López‐Doménech等人此前的研究，并做了小幅修改[1]。 下文将简要介绍该脚本的功能（脚本、演示数据及预期输出文件均已随附）。 输入文件为包含细胞核、肌动蛋白网络与线粒体三个通道的单细胞共聚焦Z-stack图像，首先对细胞核进行分割与轮廓勾勒，以获取首个同心壳层。随后，在半径依次递增1微米的30个同心壳层中，分别量化分割后的肌动蛋白网络或线粒体的面积。输出数据将保存为.csv文件，同时附带一张概览图像，用于复核分割操作是否成功。 [1] López-Doménech G, Covill-Cooke C, Ivankovic D, Halff EF, Sheehan DF, Norkett R, Birsa N, Kittler JT. Miro蛋白调控依赖微管与肌动蛋白的线粒体运输与分布. 欧洲分子生物学组织杂志（EMBO J）. 2018年2月1日;37(3):321-336. DOI: 10.15252/embj.201696380. 2018年1月8日在线发表. PMID: 29311115; PMCID: PMC5793800.