Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy
Abstract Taking advantage of the good mechanical strength of expanded Drosophila brains and to tackle their relatively large size that can complicate imaging, we apply potassium (poly)acrylate-based hydrogels for expansion microscopy (ExM), resulting in a 40x plus increased resolution of transgenic...
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Nature Portfolio
2024-12-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55305-8 |
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| author | Xuejiao Tian Tzu-Yang Lin Po-Ting Lin Min-Ju Tsai Hsin Chen Wen-Jie Chen Chia-Ming Lee Chiao-Hui Tu Jui-Cheng Hsu Tung-Han Hsieh Yi-Chung Tung Chien-Kai Wang Suewei Lin Li-An Chu Fan-Gang Tseng Yi-Ping Hsueh Chi-Hon Lee Peilin Chen Bi-Chang Chen |
| author_facet | Xuejiao Tian Tzu-Yang Lin Po-Ting Lin Min-Ju Tsai Hsin Chen Wen-Jie Chen Chia-Ming Lee Chiao-Hui Tu Jui-Cheng Hsu Tung-Han Hsieh Yi-Chung Tung Chien-Kai Wang Suewei Lin Li-An Chu Fan-Gang Tseng Yi-Ping Hsueh Chi-Hon Lee Peilin Chen Bi-Chang Chen |
| author_sort | Xuejiao Tian |
| collection | DOAJ |
| description | Abstract Taking advantage of the good mechanical strength of expanded Drosophila brains and to tackle their relatively large size that can complicate imaging, we apply potassium (poly)acrylate-based hydrogels for expansion microscopy (ExM), resulting in a 40x plus increased resolution of transgenic fluorescent proteins preserved by glutaraldehyde fixation in the nervous system. Large-volume ExM is realized by using an axicon-based Bessel lightsheet microscope, featuring gentle multi-color fluorophore excitation and intrinsic optical sectioning capability, enabling visualization of Tm5a neurites and L3 lamina neurons with photoreceptors in the optic lobe. We also image nanometer-sized dopaminergic neurons across the same intact iteratively expanded Drosophila brain, enabling us to measure the 3D expansion ratio. Here we show that at a tile scanning speed of ~1 min/mm3 with 1012 pixels over 14 hours, we image the centimeter-sized fly brain at an effective resolution comparable to electron microscopy, allowing us to visualize mitochondria within presynaptic compartments and Bruchpilot (Brp) scaffold proteins distributed in the central complex, enabling robust analyses of neurobiological topics. |
| format | Article |
| id | doaj-art-63f18951a94544908ea2cafbb9d60043 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-63f18951a94544908ea2cafbb9d600432025-01-05T12:34:59ZengNature PortfolioNature Communications2041-17232024-12-0115111610.1038/s41467-024-55305-8Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopyXuejiao Tian0Tzu-Yang Lin1Po-Ting Lin2Min-Ju Tsai3Hsin Chen4Wen-Jie Chen5Chia-Ming Lee6Chiao-Hui Tu7Jui-Cheng Hsu8Tung-Han Hsieh9Yi-Chung Tung10Chien-Kai Wang11Suewei Lin12Li-An Chu13Fan-Gang Tseng14Yi-Ping Hsueh15Chi-Hon Lee16Peilin Chen17Bi-Chang Chen18Research Center for Applied Sciences, Academia SinicaInstitute of Cellular and Organismic Biology, Academia SinicaResearch Center for Applied Sciences, Academia SinicaResearch Center for Applied Sciences, Academia SinicaResearch Center for Applied Sciences, Academia SinicaTaiwan International Graduate Program in Interdisciplinary Neuroscience, National Cheng Kung University and Academia SinicaResearch Center for Applied Sciences, Academia SinicaResearch Center for Applied Sciences, Academia SinicaResearch Center for Applied Sciences, Academia SinicaResearch Center for Applied Sciences, Academia SinicaResearch Center for Applied Sciences, Academia SinicaDepartment of Mechanical Engineering, National Taiwan UniversityInstitute of Molecular Biology, Academia SinicaDepartment of Biomedical Engineering and Environmental Sciences, National Tsing Hua UniversityResearch Center for Applied Sciences, Academia SinicaInstitute of Molecular Biology, Academia SinicaInstitute of Cellular and Organismic Biology, Academia SinicaResearch Center for Applied Sciences, Academia SinicaResearch Center for Applied Sciences, Academia SinicaAbstract Taking advantage of the good mechanical strength of expanded Drosophila brains and to tackle their relatively large size that can complicate imaging, we apply potassium (poly)acrylate-based hydrogels for expansion microscopy (ExM), resulting in a 40x plus increased resolution of transgenic fluorescent proteins preserved by glutaraldehyde fixation in the nervous system. Large-volume ExM is realized by using an axicon-based Bessel lightsheet microscope, featuring gentle multi-color fluorophore excitation and intrinsic optical sectioning capability, enabling visualization of Tm5a neurites and L3 lamina neurons with photoreceptors in the optic lobe. We also image nanometer-sized dopaminergic neurons across the same intact iteratively expanded Drosophila brain, enabling us to measure the 3D expansion ratio. Here we show that at a tile scanning speed of ~1 min/mm3 with 1012 pixels over 14 hours, we image the centimeter-sized fly brain at an effective resolution comparable to electron microscopy, allowing us to visualize mitochondria within presynaptic compartments and Bruchpilot (Brp) scaffold proteins distributed in the central complex, enabling robust analyses of neurobiological topics.https://doi.org/10.1038/s41467-024-55305-8 |
| spellingShingle | Xuejiao Tian Tzu-Yang Lin Po-Ting Lin Min-Ju Tsai Hsin Chen Wen-Jie Chen Chia-Ming Lee Chiao-Hui Tu Jui-Cheng Hsu Tung-Han Hsieh Yi-Chung Tung Chien-Kai Wang Suewei Lin Li-An Chu Fan-Gang Tseng Yi-Ping Hsueh Chi-Hon Lee Peilin Chen Bi-Chang Chen Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy Nature Communications |
| title | Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy |
| title_full | Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy |
| title_fullStr | Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy |
| title_full_unstemmed | Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy |
| title_short | Rapid lightsheet fluorescence imaging of whole Drosophila brains at nanoscale resolution by potassium acrylate-based expansion microscopy |
| title_sort | rapid lightsheet fluorescence imaging of whole drosophila brains at nanoscale resolution by potassium acrylate based expansion microscopy |
| url | https://doi.org/10.1038/s41467-024-55305-8 |
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