Low-temperature resin embedding of the whole brain for various precise structures dissection
Summary: Resin embedding combined with ultra-thin sectioning has been widely used in microscopic and electron imaging to acquire precise structural information of biological tissues. However, the existing embedding method was detrimental to quenchable fluorescent signals of precise structures and pH...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-05-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004223007824 |
| _version_ | 1797832719588130816 |
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| author | Jiaojiao Tian Yingying Chen Tao Jiang Xueyan Jia Hui Gong Xiangning Li |
| author_facet | Jiaojiao Tian Yingying Chen Tao Jiang Xueyan Jia Hui Gong Xiangning Li |
| author_sort | Jiaojiao Tian |
| collection | DOAJ |
| description | Summary: Resin embedding combined with ultra-thin sectioning has been widely used in microscopic and electron imaging to acquire precise structural information of biological tissues. However, the existing embedding method was detrimental to quenchable fluorescent signals of precise structures and pH-insensitive fluorescent dyes. Here, we developed a low-temperature chemical polymerization method named HM20-T to maintain weak signals of various precise structures and to decrease background fluorescence. The fluorescence preservation ratio of green fluorescent protein (GFP) tagged presynaptic elements and tdTomato labeled axons doubled. The HM20-T method was suitable for a variety of fluorescent dyes, such as DyLight 488 conjugated Lycopersicon esculentum lectin. Moreover, the brains also retained immunoreactivity after embedding. In summary, the HM20-T method was suitable for the characterization of multi-color labeled precise structures, which would contribute to the acquisition of complete morphology of various biological tissues and to the investigation of composition and circuit connection in the whole brain. |
| first_indexed | 2024-04-09T14:12:20Z |
| format | Article |
| id | doaj.art-bf4232fcabc843949a5b3581458c584a |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-04-09T14:12:20Z |
| publishDate | 2023-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-bf4232fcabc843949a5b3581458c584a2023-05-06T04:38:44ZengElsevieriScience2589-00422023-05-01265106705Low-temperature resin embedding of the whole brain for various precise structures dissectionJiaojiao Tian0Yingying Chen1Tao Jiang2Xueyan Jia3Hui Gong4Xiangning Li5Britton Chance Center and MoE Key Laboratory for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China; Department of Clinical Laboratory, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, ChinaBritton Chance Center and MoE Key Laboratory for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, ChinaResearch Unit of Multimodal Cross Scale Neural Signal Detection and Imaging, Chinese Academy of Medical Sciences, HUST-Suzhou Institute for Brainmatics, JITRI, Suzhou 215125, ChinaResearch Unit of Multimodal Cross Scale Neural Signal Detection and Imaging, Chinese Academy of Medical Sciences, HUST-Suzhou Institute for Brainmatics, JITRI, Suzhou 215125, ChinaBritton Chance Center and MoE Key Laboratory for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China; Research Unit of Multimodal Cross Scale Neural Signal Detection and Imaging, Chinese Academy of Medical Sciences, HUST-Suzhou Institute for Brainmatics, JITRI, Suzhou 215125, ChinaBritton Chance Center and MoE Key Laboratory for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China; Key Laboratory of Biomedical Engineering of Hainan Province, School of Biomedical Engineering, Hainan University, Haikou 570228, China; Research Unit of Multimodal Cross Scale Neural Signal Detection and Imaging, Chinese Academy of Medical Sciences, HUST-Suzhou Institute for Brainmatics, JITRI, Suzhou 215125, China; Corresponding authorSummary: Resin embedding combined with ultra-thin sectioning has been widely used in microscopic and electron imaging to acquire precise structural information of biological tissues. However, the existing embedding method was detrimental to quenchable fluorescent signals of precise structures and pH-insensitive fluorescent dyes. Here, we developed a low-temperature chemical polymerization method named HM20-T to maintain weak signals of various precise structures and to decrease background fluorescence. The fluorescence preservation ratio of green fluorescent protein (GFP) tagged presynaptic elements and tdTomato labeled axons doubled. The HM20-T method was suitable for a variety of fluorescent dyes, such as DyLight 488 conjugated Lycopersicon esculentum lectin. Moreover, the brains also retained immunoreactivity after embedding. In summary, the HM20-T method was suitable for the characterization of multi-color labeled precise structures, which would contribute to the acquisition of complete morphology of various biological tissues and to the investigation of composition and circuit connection in the whole brain.http://www.sciencedirect.com/science/article/pii/S2589004223007824NeuroscienceTechniques in neuroscienceBiological sciences research methodologiesMethodology in biological sciences |
| spellingShingle | Jiaojiao Tian Yingying Chen Tao Jiang Xueyan Jia Hui Gong Xiangning Li Low-temperature resin embedding of the whole brain for various precise structures dissection iScience Neuroscience Techniques in neuroscience Biological sciences research methodologies Methodology in biological sciences |
| title | Low-temperature resin embedding of the whole brain for various precise structures dissection |
| title_full | Low-temperature resin embedding of the whole brain for various precise structures dissection |
| title_fullStr | Low-temperature resin embedding of the whole brain for various precise structures dissection |
| title_full_unstemmed | Low-temperature resin embedding of the whole brain for various precise structures dissection |
| title_short | Low-temperature resin embedding of the whole brain for various precise structures dissection |
| title_sort | low temperature resin embedding of the whole brain for various precise structures dissection |
| topic | Neuroscience Techniques in neuroscience Biological sciences research methodologies Methodology in biological sciences |
| url | http://www.sciencedirect.com/science/article/pii/S2589004223007824 |
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