Correlative light and electron microscopy for complex cellular structures on PDMS substrates with coded micro-patterns
文献类型:期刊论文
作者 | Sun, Rong1,2,3; Chen, Xi4; Yin, Chun-Ying1,2,3; Qi, Lei2,3; Lau, Pak-Ming2,3; Han, Hua4; Bi, Guo-Qiang1,2,3,5 |
刊名 | LAB ON A CHIP |
出版日期 | 2018-12-21 |
卷号 | 18期号:24页码:3840-3848 |
ISSN号 | 1473-0197 |
关键词 | CLEM PDMS substrates |
DOI | 10.1039/c8lc00703a |
英文摘要 | Fluorescence light microscopy (FLM) is commonly used for localizing specific cellular and subcellular targets. Electron microscopy (EM), on the other hand, can reveal ultrastructural details of cellular architectures beyond the limit of optical resolution. Correlative light and electron microscopy (CLEM) that combines the two techniques has proven valuable in various cell biological applications that require both specificity and resolution. Here, we report an efficient and easy-to-use CLEM system, and its applications in studying neuronal synapses. The system utilizes patterned symbols to encode coordinates on micro-fabricated polydimethylsiloxane (PDMS) substrates, on which dissociated primary hippocampal neurons grow and form synaptic connections. After imaging and localizing specifically labeled synapses with FLM, samples are embedded in resin blocks and sectioned for EM analysis. The patterned symbols on PDMS substrates provide coordinate information, allowing efficient co-registration between FLM and EM images with high precision. A custom-developed software package achieves automated EM image collection, FLM/EM alignment, and EM navigation. With this CLEM system, we have obtained high quality electron tomograms of fluorescently labeled synapses along dendrites of hippocampal neurons and analyzed docking statistics of synaptic vesicles (SVs) in different subtypes of excitatory synapses. This technique provides an efficient approach to combine functional studies with ultrastructural analysis of heterogeneous neuronal synapses, as well as other subcellular structures in general. |
WOS关键词 | FLUORESCENCE ; TOMOGRAPHY ; ORGANIZATION ; ENDOCYTOSIS ; MULTISCALE ; SYNAPSES ; NEURONS ; CULTURE ; CELLS |
资助项目 | National Key R&D Program of China[2016YFA0400900] ; CAS[XDB02050000] ; National Natural Science Foundation of China[31630030] |
WOS研究方向 | Biochemistry & Molecular Biology ; Chemistry ; Science & Technology - Other Topics |
语种 | 英语 |
出版者 | ROYAL SOC CHEMISTRY |
WOS记录号 | WOS:000452320500011 |
源URL | [http://ir.ia.ac.cn/handle/173211/25681] |
专题 | 类脑智能研究中心_微观重建与智能分析 |
通讯作者 | Chen, Xi; Bi, Guo-Qiang |
作者单位 | 1.Univ Sci & Technol China, Ctr Integrat Imaging, Natl Lab Phys Sci Microscale, Hefei, Anhui, Peoples R China 2.Univ Sci & Technol China, CAS Key Lab Brain Funct & Dis, 443 Huangshan Rd, Hefei 230027, Anhui, Peoples R China 3.Univ Sci & Technol China, Sch Life Sci, 443 Huangshan Rd, Hefei 230027, Anhui, Peoples R China 4.Chinese Acad Sci, Inst Automat, 95 Zhongguancun East Rd, Beijing 100190, Peoples R China 5.Univ Sci & Technol China, CAS Ctr Excellence Brain Sci & Intelligence Techn, Hefei, Anhui, Peoples R China |
推荐引用方式 GB/T 7714 | Sun, Rong,Chen, Xi,Yin, Chun-Ying,et al. Correlative light and electron microscopy for complex cellular structures on PDMS substrates with coded micro-patterns[J]. LAB ON A CHIP,2018,18(24):3840-3848. |
APA | Sun, Rong.,Chen, Xi.,Yin, Chun-Ying.,Qi, Lei.,Lau, Pak-Ming.,...&Bi, Guo-Qiang.(2018).Correlative light and electron microscopy for complex cellular structures on PDMS substrates with coded micro-patterns.LAB ON A CHIP,18(24),3840-3848. |
MLA | Sun, Rong,et al."Correlative light and electron microscopy for complex cellular structures on PDMS substrates with coded micro-patterns".LAB ON A CHIP 18.24(2018):3840-3848. |
入库方式: OAI收割
来源:自动化研究所
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