Assembly and operation of an imaging system for long-term monitoring of bioluminescent and fluorescent reporters in plants.

Abstract

Background Non-invasive reporter systems are powerful tools to query physiological and transcriptional responses in organisms. For example, fluorescent and bioluminescent reporters have revolutionized cellular and organismal assays and have been used to study plant responses to abiotic and biotic stressors. Integrated, cooled charge-coupled device (CCD) camera systems have been developed to image bioluminescent and fluorescent signals in a variety of organisms; however, these integrated long-term imaging systems are expensive. Results We have developed self-assembled systems for both growing and monitoring plant fluorescence and bioluminescence for long-term experiments under controlled environmental conditions. This system combines environmental growth chambers with high-sensitivity CCD cameras, multi-wavelength LEDs, open-source software, and several options for coordinating lights with imaging. This easy-to-assemble system can be used for short and long-term imaging of bioluminescent reporters, acute light-response, circadian rhythms, delayed fluorescence, and fluorescent-protein-based assays in vivo. Conclusions We have developed two self-assembled imaging systems that will be useful to researchers interested in continuously monitoring in vivo reporter systems in various plant species.

Plant phenotyping relevance

植物の蛍光・生物発光を長期的に画像取得する自作システムを開発しており、植物の生理状態・レポーター応答の非侵襲的測定が中心的な方法論的貢献である。

Code and data availability

The paper's authors explicitly state code availability for two public GitHub repositories: the LightsCameraAction µManager plug-in for coordinating lights with imaging (System 1) and the GreenhamLab/CCD_Imaging repository containing Python scripts for time-series acquisition and light scheduling plus sample data for bi