Automating measurements of fluorescent signals in freely moving plant leaf specimens
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- Kinoshita Natsuko
- Faculty of Life and Environmental Sciences, University of Tsukuba
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- Sugita Aki
- Faculty of Life and Environmental Sciences, University of Tsukuba Department of Evolutionary Studies of Biosystems, School of Advanced Sciences, SOKENDAI (The Graduate University for Advanced Studies)
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- Lustig Barry
- Cormorant Group
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- Betsuyaku Shigeyuki
- Faculty of Life and Environmental Sciences, University of Tsukuba
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- Morishita Tatsuji
- Leica Microsystems
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Abstract
<p>Existing methods to quantify fluorescent signals are primarily limited to non-moving objects or tracking a limited number of cells. These techniques, however, are unsuitable for measuring fluorescent signals in time-lapse experiments using plant specimens that move naturally during a course of imaging. We developed an automated method to measure fluorescent signal intensities in transgenic Arabidopsis plants using a stereomicroscope with standard microscopy software. The features of our technique include: 1) recognizing the shape of plant specimens using autofluorescent signals; 2) merging targeted fluorescent signals to specimen outlines; 3) extracting signals within the shape of specimens from their background signals. Our method facilitates the measurement of fluorescent signals on freely moving plant leaves that are physically unrestrained. The method we developed addresses the challenge of recognizing plant shapes without relying on: a) manual definition which is prone to subjectivity and human error; b) introducing stable fluorescent markers to define plant shapes; c) recognizing plant shapes from bright field images which include a wide range of colors and background noise; d) unnecessarily stressing plants by immobilizing them; e) the use of multiple software packages or software development expertise.</p>
Journal
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- Plant Biotechnology
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Plant Biotechnology 36 (1), 7-11, 2019-03-25
Japanese Society for Plant Biotechnology
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Details 詳細情報について
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- CRID
- 1390564238095541504
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- NII Article ID
- 130007628753
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- NII Book ID
- AA11250821
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- ISSN
- 13476114
- 13424580
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- NDL BIB ID
- 030207447
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
- KAKEN
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- Abstract License Flag
- Disallowed