Low-cost calcium fluorometry for long-term nanoparticle studies in living cells

Connor L Beck,Anja Kunze,Clark J Hickman

doi:10.1038/s41598-020-69412-1

Connor L Beck, Anja Kunze + Show 1 more

Open Access

https://doi.org/10.1038/s41598-020-69412-1

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Abstract

Calcium fluorometry is critical to determine cell homeostasis or to reveal communication patterns in neuronal networks. Recently, characterizing calcium signalling in neurons related to interactions with nanomaterials has become of interest due to its therapeutic potential. However, imaging of neuronal cell activity under stable physiological conditions can be either very expensive or limited in its long-term capability. Here, we present a low-cost, portable imaging system for long-term, fast-scale calcium fluorometry in neurons. Using the imaging system, we revealed temperature-dependent changes in long-term calcium signalling in kidney cells and primary cortical neurons. Furthermore, we introduce fast-scale monitoring of synchronous calcium activity in neuronal cultures in response to nanomaterials. Through graph network analysis, we found that calcium dynamics in neurons are temperature-dependent when exposed to chitosan-coated nanoparticles. These results give new insights into nanomaterial-interaction in living cultures and tissues based on calcium fluorometry and graph network analysis.

Highlights

Calcium fluorometry is critical to determine cell homeostasis or to reveal communication patterns in neuronal networks
The first set demonstrates the capability of long-term image acquisition through monitoring temperature-dependent calcium dynamics in HEK293 cells in a lab-extern cell culture facility and primary cortical neurons grown in our lab
We were able to characterize slow, long-term calcium dynamics in primary cortical neuron cultures based on (a) temperature-dependent temporal changes in calcium signalling, (b) calcium events associated with cell death, and (c) fast-scale spatiotemporal changes in synchronous calcium dynamics associated with the uptake of chitosan-coated nanoparticles

Summary

Introduction

Calcium fluorometry is critical to determine cell homeostasis or to reveal communication patterns in neuronal networks. We were able to characterize slow, long-term calcium dynamics in primary cortical neuron cultures based on (a) temperature-dependent temporal changes in calcium signalling, (b) calcium events associated with cell death, and (c) fast-scale spatiotemporal changes in synchronous calcium dynamics associated with the uptake of chitosan-coated nanoparticles. This low-cost, portable, and easy to assemble long-term imaging platform can expand fluorescent imaging of neuronal cell dynamics to low-resource environments, field settings, and even classrooms. It has the potential to expand knowledgegaining and next-generation neuro-tool development to a broader academic spectrum

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