A modular in-incubator microscope for longitudinal live-cell microscopy

Abstract

Longitudinal live-cell imaging is valuable for characterizing morphological and phenotypic changes in biological systems. However, conventional approaches rely on manual microscope operation, which is labor-intensive, limits imaging frequency, and disrupts cellular environments. These constraints increase experimental variability and restrict both the duration and temporal resolution of continuous imaging. Although automated microscopes partially address these limitations, existing solutions are often constrained by the cost, footprint, and inflexibility of in-incubator microscopes or stage-top incubators. Here, we present an automated in-incubator epifluorescence microscope designed for extended operation. The system features a modular architecture with optional automated multi-fluorescence imaging, plate scanning, and compatibility with fluidic automation devices. By placing the light sources and control electronics outside the incubator, the platform improves thermal stability and long-term operational reliability. This approach enables continuous, high-frequency imaging over extended durations, providing a source of rich data for quantifying time-dependent tissue phenotypes, morphological remodeling, and transient biological processes.