Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using Fluorescein
Plants use CO2, water, and light energy to generate carbohydrates through photosynthesis. During daytime, these carbohydrates are polymerized, leading to the accumulation of starch granules in chloroplasts. The catabolites produced by the degradation of these chloroplast starch granules are used for...
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Bio-protocol LLC
2024-11-01
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| Online Access: | https://bio-protocol.org/en/bpdetail?id=5103&type=0 |
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| author | Shintaro Ichikawa Yutaka Kodama |
| author_facet | Shintaro Ichikawa Yutaka Kodama |
| author_sort | Shintaro Ichikawa |
| collection | DOAJ |
| description | Plants use CO2, water, and light energy to generate carbohydrates through photosynthesis. During daytime, these carbohydrates are polymerized, leading to the accumulation of starch granules in chloroplasts. The catabolites produced by the degradation of these chloroplast starch granules are used for physiological responses and plant growth. Various staining methods, such as iodine staining, have previously been used to visualize the accumulation of chloroplast starch granules; however, these staining methods cannot be used to image live cells and/or provide confocal images with non-specific signals. In this study, we developed a new imaging method for the fluorescent observation of chloroplast starch granules in living plant cells by staining with fluorescein, a widely available fluorescent dye. This simple staining method, which involves soaking a leaf disk in staining solution, shows high specificity in confocal images. Fluorescent images of the stained tissue allow the cellular starch content of living cells to be quantified with the same level of accuracy as a conventional biochemical method (amyloglucosidase/α-amylase method). Fluorescein staining thus not only enables the easy and clear observation of chloroplast starch granules but also allows for precise quantification in living cells. |
| format | Article |
| id | doaj-art-654d4a42a1a24ed19a6ebdde715024d5 |
| institution | OA Journals |
| issn | 2331-8325 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Bio-protocol LLC |
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| series | Bio-Protocol |
| spelling | doaj-art-654d4a42a1a24ed19a6ebdde715024d52025-08-20T01:57:20ZengBio-protocol LLCBio-Protocol2331-83252024-11-01142110.21769/BioProtoc.5103Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using FluoresceinShintaro Ichikawa0Yutaka Kodama1Center for Bioscience Research and Education, Utsunomiya University, Tochigi, JapanGraduate School of Regional Development and Creativity, Utsunomiya University, Tochigi, JapanCenter for Bioscience Research and Education, Utsunomiya University, Tochigi, JapanGraduate School of Regional Development and Creativity, Utsunomiya University, Tochigi, JapanPlants use CO2, water, and light energy to generate carbohydrates through photosynthesis. During daytime, these carbohydrates are polymerized, leading to the accumulation of starch granules in chloroplasts. The catabolites produced by the degradation of these chloroplast starch granules are used for physiological responses and plant growth. Various staining methods, such as iodine staining, have previously been used to visualize the accumulation of chloroplast starch granules; however, these staining methods cannot be used to image live cells and/or provide confocal images with non-specific signals. In this study, we developed a new imaging method for the fluorescent observation of chloroplast starch granules in living plant cells by staining with fluorescein, a widely available fluorescent dye. This simple staining method, which involves soaking a leaf disk in staining solution, shows high specificity in confocal images. Fluorescent images of the stained tissue allow the cellular starch content of living cells to be quantified with the same level of accuracy as a conventional biochemical method (amyloglucosidase/α-amylase method). Fluorescein staining thus not only enables the easy and clear observation of chloroplast starch granules but also allows for precise quantification in living cells.https://bio-protocol.org/en/bpdetail?id=5103&type=0 |
| spellingShingle | Shintaro Ichikawa Yutaka Kodama Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using Fluorescein Bio-Protocol |
| title | Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using Fluorescein |
| title_full | Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using Fluorescein |
| title_fullStr | Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using Fluorescein |
| title_full_unstemmed | Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using Fluorescein |
| title_short | Fluorescent Staining and Quantification of Starch Granules in Chloroplasts of Live Plant Cells Using Fluorescein |
| title_sort | fluorescent staining and quantification of starch granules in chloroplasts of live plant cells using fluorescein |
| url | https://bio-protocol.org/en/bpdetail?id=5103&type=0 |
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