ALBERT

Description: © The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Lagache, T., Hanson, A., Perez-Ortega, J. E., Fairhall, A., & Yuste, R. Tracking calcium dynamics from individual neurons in behaving animals. Plos Computational Biology, 17(10), (2021): e1009432, https://doi.org/10.1371/journal.pcbi.1009432.

Description: Measuring the activity of neuronal populations with calcium imaging can capture emergent functional properties of neuronal circuits with single cell resolution. However, the motion of freely behaving animals, together with the intermittent detectability of calcium sensors, can hinder automatic monitoring of neuronal activity and their subsequent functional characterization. We report the development and open-source implementation of a multi-step cellular tracking algorithm (Elastic Motion Correction and Concatenation or EMC2) that compensates for the intermittent disappearance of moving neurons by integrating local deformation information from detectable neurons. We demonstrate the accuracy and versatility of our algorithm using calcium imaging data from two-photon volumetric microscopy in visual cortex of awake mice, and from confocal microscopy in behaving Hydra, which experiences major body deformation during its contractions. We quantify the performance of our algorithm using ground truth manual tracking of neurons, along with synthetic time-lapse sequences, covering a wide range of particle motions and detectability parameters. As a demonstration of the utility of the algorithm, we monitor for several days calcium activity of the same neurons in layer 2/3 of mouse visual cortex in vivo, finding significant turnover within the active neurons across days, with only few neurons that remained active across days. Also, combining automatic tracking of single neuron activity with statistical clustering, we characterize and map neuronal ensembles in behaving Hydra, finding three major non-overlapping ensembles of neurons (CB, RP1 and RP2) whose activity correlates with contractions and elongations. Our results show that the EMC2 algorithm can be used as a robust and versatile platform for neuronal tracking in behaving animals.

Description: R.Y. was supported by the NSF (CRCNS 1822550), the NEI (R01EY011787), the NIMH (R01MH115900), and Vannevar Bush Faculty Award (ONR N000142012828). T.L. was supported by the Fondation pour la Recherche Médicale (https://www.frm.org/) and the Philippe Foundation (https://www.philippefoundation.org/). A.H. was supported by the NIMH (T32MH018870). J.P.-O. was supported by the CONACYT (CVU365863). ALF was supported by NSF (CRCNS 1822550), the Simons Foundation Collaboration for the Global Brain (542975SPI) and the Weill NeuroHub (https://www.weillneurohub.org/).