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The cellular distribution of receptors, ligands, and activation state must be integrated with information about the cellular distribution of metabolites in relation to metabolic fluxes and signaling dynamics in order to achieve the promise of in vivo biochemistry. Genetically encoded sensors are engineered fluorescent proteins that have been developed for a wide range of small molecules, such as ions and metabolites, or to report biophysical processes, such as transmembrane voltage or tension. First steps have been taken to monitor the activity of transporters in vivo. Advancements in imaging technologies and specimen handling and stimulation have enabled researchers in plant sciences to implement sensor technologies in intact plants. Here, we provide a brief history of the development of genetically encoded sensors and an overview of the types of sensors available for quantifying and visualizing ion and metabolite distribution and dynamics. We further discuss the pros and cons of specific sensor designs, imaging systems, and sample manipulations, provide advice on the choice of technology, and give an outlook into future developments.<\/jats:p>","DOI":"10.1093\/plphys\/kiab353","type":"journal-article","created":{"date-parts":[[2021,7,23]],"date-time":"2021-07-23T11:13:33Z","timestamp":1627038813000},"page":"485-503","source":"Crossref","is-referenced-by-count":57,"title":["Designs, applications, and limitations of genetically encoded fluorescent sensors to explore plant biology"],"prefix":"10.1093","volume":"187","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0217-6731","authenticated-orcid":false,"given":"Mayuri","family":"Sadoine","sequence":"first","affiliation":[{"name":"Molecular Physiology, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"}]},{"given":"Yuuma","family":"Ishikawa","sequence":"additional","affiliation":[{"name":"Molecular Physiology, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"},{"name":"Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Chikusa, Nagoya 464-8601, Japan"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2568-7323","authenticated-orcid":false,"given":"Thomas J.","family":"Kleist","sequence":"additional","affiliation":[{"name":"Molecular Physiology, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4332-369X","authenticated-orcid":false,"given":"Michael M.","family":"Wudick","sequence":"additional","affiliation":[{"name":"Molecular Physiology, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"},{"name":"Cluster of Excellence on Plant Sciences, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7107-4942","authenticated-orcid":false,"given":"Masayoshi","family":"Nakamura","sequence":"additional","affiliation":[{"name":"Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Chikusa, Nagoya 464-8601, Japan"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7529-9244","authenticated-orcid":false,"given":"Guido","family":"Grossmann","sequence":"additional","affiliation":[{"name":"Cluster of Excellence on Plant Sciences, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"},{"name":"Institute for Cell and Interaction Biology, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6465-0115","authenticated-orcid":false,"given":"Wolf B.","family":"Frommer","sequence":"additional","affiliation":[{"name":"Molecular Physiology, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"},{"name":"Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Chikusa, Nagoya 464-8601, Japan"},{"name":"Cluster of Excellence on Plant Sciences, Heinrich-Heine-University D\u00fcsseldorf, D\u00fcsseldorf 40225, Germany"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0895-1914","authenticated-orcid":false,"given":"Cheng-Hsun","family":"Ho","sequence":"additional","affiliation":[{"name":"Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan"}]}],"member":"286","published-online":{"date-parts":[[2021,8,30]]},"reference":[{"key":"2022042006321823700_kiab353-B1","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1016\/j.cub.2006.03.035","article-title":"DRB4-dependent TAS3 trans-acting siRNAs control leaf morphology through AGO7","volume":"16","author":"Adenot","year":"2006","journal-title":"Curr Biol"},{"key":"2022042006321823700_kiab353-B2","doi-asserted-by":"crossref","first-page":"6455","DOI":"10.1074\/jbc.M807657200","article-title":"Crystal structures of 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