BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

Michael Zabolocki, Kasandra McCormack, Mark van den Hurk, Bridget Milky, Andrew P. Shoubridge, Robert Adams, Jenne Tran, Anita Mahadevan-Jansen, Philipp Reineck, Jacob Thomas, Mark R. Hutchinson, Carmen K.H. Mak, Adam Añonuevo, Leon H. Chew, Adam J. Hirst, Vivian M. Lee, Erin Knock, Cedric Bardy

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article number5550
Number of pages19
JournalNature Communications
Volume11
Issue number1
DOIs
Publication statusPublished - 3 Nov 2020

Keywords

  • Fluorescence imaging
  • Induced pluripotent stem cells
  • Neurophysiology
  • Optogenetics
  • Patch clamp

Fingerprint Dive into the research topics of 'BrainPhys neuronal medium optimized for imaging and optogenetics in vitro'. Together they form a unique fingerprint.

Cite this