Sholl analysis of mitochondria from article The adaptor protein Miro1 modulates horizontal transfer of mitochondria in cancer
Description
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.
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Steps to reproduce
For sample preparation details, refer to Novak & Nahacka et al., The adaptor protein Miro1 modulates horizontal transfer of mitochondria in mouse melanoma models, published in Cell Reports.