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. 2019 May 29;7:87. doi: 10.3389/fcell.2019.00087

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Copyright © 2019 Song, Laureano, Patel, Yip, Jadali and Kwan.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Sparse labeling of iMOP-derived neurons for pseudotemporal ordering. (A) Nuclear labeling with Hoechst and immunostaining of iMOP-derived neuron with antibodies against CDKN1B and TUBB3. (B) Expression of GFP in TUBB3 marked cells by transfecting a GFP overexpression plasmid for sparse labeling and neurite tracing. Arrowheads mark the path of the neurite in a single neuron. (C) Workflow to identify GFP+ TUBB3– and GFP+ TUBB3+ cells for fluoresence intensity and neurite length measurements. (D) Arrangement of iMOP-derived neurons based on TUBB3 labeling and neurite lengths to generate a pseudotemporal ordering of cells that represent a differentiation trajectory. Scale bars as noted.