Design and validation of an open-source modular Microplate Photoirradiation System for high-throughput photobiology...

Area of Science:

• Photobiology
• Biophotonics
• Cellular Assays

Background:

• Current photobiology research is hindered by the lack of high-throughput, precise, and tunable irradiation devices.
• Existing solutions are often expensive, difficult to replicate, and limit experimental standardization and scalability.
• A need exists for flexible, open-source instrumentation for advanced photobiology studies.

Purpose of the Study:

• To develop and validate an open-source Microplate Photoirradiation System (MPS) for high-throughput cell irradiation.
• To enable precise control over light parameters (irradiance, duration, pulsation) in standard microplate formats.
• To facilitate reproducible and scalable experiments in photobiology research.

Main Methods:

• Development of an open-source MPS with independently controlled LEDs, Wi-Fi connectivity, and a programmable GUI.
• Implementation of web-based GUI for generating light program files and wireless parameter uploading.
• Validation using a porphyrin metabolism assay in fibroblasts, inducing Protoporphyrin IX (PpIX) fluorescence with 5-aminolevulinic acid (ALA) and irradiating with 405nm light.

Main Results:

• The MPS successfully delivered precise irradiance and timing for photoirradiation experiments in 96 and 24-well microplates.
• PpIX photobleaching kinetics were measured in live, adherent cells, demonstrating system reliability and comparability across formats.
• High-throughput capability allowed fitting fluorescence data to a bioexponential decay model and revealed photobleaching dependency on duty-cycle, not frequency.

Conclusions:

• The open-source Microplate Photoirradiation System (MPS) effectively addresses the need for high-throughput, precise cell irradiation in photobiology.
• The system enhances experimental standardization, reproducibility, and scalability for diverse photobiology applications.
• The MPS facilitates detailed kinetic studies and mechanistic investigations of light-cell interactions.