# LUMIN: an automated graphical analysis toolbox for high-throughput calcium imaging of in vitro neuronal cultures

## TL;DR

LUMIN is a new software tool for analyzing calcium imaging data from lab-grown neurons, enabling efficient and accurate study of neuronal activity for drug screening and disease modeling.

## Contribution

LUMIN introduces a scalable, automated graphical analysis tool specifically optimized for high-throughput calcium imaging of in vitro neuronal cultures.

## Key findings

- LUMIN processes thousands of cells within hours on a standard laptop due to its efficient data processing and linear time complexity.
- The software successfully demonstrated performance in analyzing human stem cell-derived ventral midbrain neurons using pharmacological compounds.
- LUMIN includes two modules for analyzing transiently active and quiescent neurons, making it broadly applicable to diverse neuronal cultures.

## Abstract

Human pluripotent stem cell-derived in vitro models, combined with advances in high-content live-cell calcium imaging, provide a powerful platform for disease modeling and drug screening. However, most calcium imaging analysis tools have been developed for in vivo applications with suboptimal features for processing data from cultured neurons. Here, we present LUMIN (Live-cell User Module for Imaging and analysis of Neuronal activity), a scalable and broadly applicable software that integrates a graphical user interface with automated data analysis encompassing single-cell segmentation, signal extraction, and activity quantification. Two analysis modules for transiently active or quiescent stimuli-evoked neurons make it broadly applicable for cultures with distinct functional properties. Its efficient data processing and linear time complexity enable processing thousands of cells within hours (~ 7000 cells from ~ 70 recordings per 30 min) on a standard laptop. We demonstrate LUMIN’s performance on human stem cell-derived ventral midbrain neurons through two applications: (1) modulation of spontaneous activity with well-characterized pharmacological compounds, and (2) evoking responses in quiescent neurons upon compound stimulation.

The online version contains supplementary material available at 10.1038/s41598-026-40269-0.