An Open-Source Pipeline for Calcium Imaging and All-Optical Physiology in Human Stem Cell-Derived Neurons

Area of Science:

• Neuroscience
• Biotechnology
• Genetics

Background:

• Understanding neuronal activity is key for brain function and neurological disease modeling.
• Current methods for profiling neuronal activity lack scalability and automation.

Purpose of the Study:

• To present an open-source, scalable platform for high-throughput, single-cell resolution profiling of neuronal activity.
• To enable functional phenotyping of human stem cell-derived neurons in disease models.

Main Methods:

• Integration of optogenetic stimulation, calcium imaging, and automated acquisition.
• Development of single-cell and network analysis tools.
• Utilized CRISPR-Cas9 to generate GCaMP6s knock-in hiPSC lines.

Main Results:

• Robust quantification of spontaneous and evoked neuronal activity in hundreds of neurons.
• Demonstrated platform versatility across CDKL5 Deficiency, SSADH Deficiency, and Tuberous Sclerosis Complex (TSC) models.
• Showcased partial pharmacological reversal in a TSC model.

Conclusions:

• The platform offers a generalizable framework for scalable functional phenotyping.
• Enables high-throughput screening of human neuronal models for drug discovery.
• Facilitates linking single-cell dynamics to network-level measures for disease insights.