Sorkin Maria L, Markham Kathleen K, Zorich Stevan, Menon Ananda, Edgeworth Kristen N, Ricono Angela, Bryant Douglas, Bart Rebecca, Nusinow Dmitri A, Greenham Kathleen
Donald Danforth Plant Science Center, St. Louis, MO, USA.
Department of Biological and Biomedical Sciences, Washington University in St. Louis, St. Louis, MO, USA.
Plant Methods. 2023 Mar 1;19(1):19. doi: 10.1186/s13007-023-00997-0.
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.
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.
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.
非侵入性报告系统是用于探究生物体生理和转录反应的强大工具。例如,荧光和生物发光报告基因彻底改变了细胞和生物体检测方法,并已用于研究植物对非生物和生物胁迫的反应。已开发出集成式冷却电荷耦合器件(CCD)相机系统,用于对多种生物体中的生物发光和荧光信号进行成像;然而,这些集成式长期成像系统价格昂贵。
我们开发了自组装系统,用于在可控环境条件下对植物荧光和生物发光进行长期实验的生长和监测。该系统将环境生长室与高灵敏度CCD相机、多波长发光二极管、开源软件以及多种用于协调光照与成像的选项相结合。这个易于组装的系统可用于生物发光报告基因的短期和长期成像、急性光反应、昼夜节律、延迟荧光以及基于荧光蛋白的体内检测。
我们开发了两种自组装成像系统,这将对有兴趣持续监测各种植物物种体内报告系统的研究人员有用。