IncuSense: IoT-Enabled Wireless Long-Term Multiparametric Cell Culture Incubator Monitoring System

Abstract Cell-culture incubators are indispensable for in vitro research and new approach methods (NAMs), yet they remain vulnerable to undetected environmental fluctuations that degrade experimental reproducibility and compromise biological sample integrity. Existing monitoring solutions partially mitigate this risk but are frequently encumbered by wired infrastructure, external signal-transmitter assemblies, or a large form factor that hinder integration with diverse workflows. This study presents IncuSense, a miniaturized, battery-powered, wireless environmental monitoring platform capable of sensing temperature, humidity, and CO 2 in real-time. The system integrates a photoacoustic NDIR sensor with a microcontroller, enabling periodic data acquisition and transmission to local or cloud-hosted dashboards for real-time visualization and automated alerts. Comprehensive thermal characterization identified a critical limitation of continuous MCU operation: Wi‑Fi standby power consumption induced progressive self-heating, yielding an internal temperature rise of approximately 18 °C and systematic bias in temperature and humidity readouts. To address this challenge, an intermittent power-cycling strategy was implemented to gate power to all ESP32 subsystems except the real-time clock, reducing the average current draw by more than 90-fold to 0.92 mA, suppressing thermal drift, and extending operational lifetime. Benchmarking against a NIST-traceable sensor demonstrated high accuracy and precision in environmental measurements, and incubator failure recording. 100-day deployment in a CO 2 incubator further confirmed long-term stability, with minimal drift and demonstrated its ability to detect transient fluctuations caused by routine incubator access. Collectively, these results establish IncuSense as a compact, cost-effective, and adaptable platform for continuous long-term deployment with periodic data acquisition, facilitating environmental standardization in biomedical and biomanufacturing workflows, and may support the standardization of NAMs.