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https://hdl.handle.net/2440/129929
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Type: | Journal article |
Title: | BrainPhys neuronal medium optimized for imaging and optogenetics in vitro |
Author: | Zabolocki, M. McCormack, K. van den Hurk, M. Milky, B. Shoubridge, A.P. Adams, R. Tran, J. Mahadevan-Jansen, A. Reineck, P. Thomas, J. Hutchinson, M.R. Mak, C.K.H. Añonuevo, A. Chew, L.H. Hirst, A.J. Lee, V.M. Knock, E. Bardy, C. |
Citation: | Nature Communications, 2020; 11(1):1-19 |
Publisher: | Nature Research |
Issue Date: | 2020 |
ISSN: | 2041-1723 2041-1723 |
Statement of Responsibility: | Michael Zabolocki, Kasandra McCormack, Mark van den Hurk, Bridget Milky, Andrew P. Shoubridge, Robert Adams ... et al. |
Abstract: | The capabilities of imaging technologies, fluorescent sensors, and optogenetics tools for cell biology are advancing. In parallel, cellular reprogramming and organoid engineering are expanding the use of human neuronal models in vitro. This creates an increasing need for tissue culture conditions better adapted to live-cell imaging. Here, we identify multiple caveats of traditional media when used for live imaging and functional assays on neuronal cultures (i.e., suboptimal fluorescence signals, phototoxicity, and unphysiological neuronal activity). To overcome these issues, we develop a neuromedium called BrainPhys™ Imaging (BPI) in which we optimize the concentrations of fluorescent and phototoxic compounds. BPI is based on the formulation of the original BrainPhys medium. We benchmark available neuronal media and show that BPI enhances fluorescence signals, reduces phototoxicity and optimally supports the electrical and synaptic activity of neurons in culture. We also show the superior capacity of BPI for optogenetics and calcium imaging of human neurons. Altogether, our study shows that BPI improves the quality of a wide range of fluorescence imaging applications with live neurons in vitro while supporting optimal neuronal viability and function. |
Keywords: | Brain Nerve Net Neurons Synapses Cells, Cultured Cerebrospinal Fluid Animals Humans Rats Culture Media Diagnostic Imaging Cell Survival Action Potentials Osmolar Concentration Light Fluorescence Induced Pluripotent Stem Cells Signal-To-Noise Ratio Optogenetics |
Rights: | © The Author(s) 2020. 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/. |
DOI: | 10.1038/s41467-020-19275-x |
Grant ID: | http://purl.org/au-research/grants/arc/CE140100003 http://purl.org/au-research/grants/arc/FT180100565 |
Published version: | http://dx.doi.org/10.1038/s41467-020-19275-x |
Appears in Collections: | Aurora harvest 8 Medicine publications |
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