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. 2018 Nov 13;5:180241. doi: 10.1038/sdata.2018.241

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Copyright © 2018, The Author(s)

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ The Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the metadata files made available in this article.

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Inducible pluripotent stem cells from human ALS patients (iPSCs) were differentiated via neuronal progenitor cells (NPCs) for final maturation to spinal motor neurons (MNs) in Zona microfluidic chambers (MFCs) to obtain compartmentalized cultures with a defined distal – to – proximal axon alignment in microchannels. Live imaging was performed at strictly standardized readout positions to obtain distal versus proximal organelle trafficking data with Mito- and Lysotracker. Resultant movies were sorted into a hierarchical folder tree to define meta conditions for automated dynamic organelle tracking and static morphology analysis in FIJI and subsequent data mining and result assembly in KNIME. Deviation from proximal Ctrl Mock was expressed as Z-score for a total of 40 parameters to obtain a HC signature as comprehensive quantitative descriptor for phenotypic deviations in diseased and drug/RNAi – manipulated states. Finally, entire signatures were ranked and pair-wisely agglomerated by hierarchical clustering.