Multicolor Single-Molecule Localization Super-Resolution Microscopy

Leiting, 潘雷霆 Pan; Fen, 胡芬 Hu; Xinzheng, 张心正 Zhang; Jingjun, 许京军 Xu

doi:10.3788/aos201737.0318010

光学学报

2017

DOI: 10.3788/aos201737.0318010

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Multicolor Single-Molecule Localization Super-Resolution Microscopy

潘雷霆 Pan Leiting¹,

胡芬 Hu Fen²,

张心正 Zhang Xinzheng³

et al.

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2024

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Multicolor single-molecule localization microscopy: review and prospect

Chen,

Wang,

Huang

et al. 2024

PhotoniX

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Single-molecule localization microscopy (SMLM) surpasses the diffraction limit by randomly switching fluorophores between fluorescent and dark states, precisely pinpointing the resulted isolated emission patterns, thereby reconstructing the super-resolution images based on the accumulated locations of thousands to millions of single molecules. This technique achieves a ten-fold improvement in resolution, unveiling the intricate details of molecular activities and structures in cells and tissues. Multicolor SMLM extends this capability by imaging distinct protein species labeled with various fluorescent probes, providing insights into structural intricacies and spatial relationships among different targets. This review explores recent advancements in multicolor SMLM, evaluates the strengths and limitations of each variant, and discusses the future prospects.

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Multicolor single-molecule localization microscopy: review and prospect

Chen,

Wang,

Huang

et al. 2024

PhotoniX

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二抗物种特异性对双色storm成像的影响（特邀）

Deng Shiyu,

Hu Fen,

Hou Mengdi

et al. 2024

激光与光电子学进展

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Stochastic optical reconstruction microscopy (STORM), a superresolution imaging technology based on immunofluorescence, has gained popularity owing to its straightforward principle, simple optical path, and excellent spatial resolution. However, the enhanced resolution demands greater specificity from antibodies. While direct labeling with primary antibodies is an option, indirect labeling using a "primary + secondary antibody" combination is more commonly employed in practical applications. Considering the issue of species specificity with secondary antibodies, preadsorption is required to increase specificity during production. In this study, we aim to explore the effect of secondary antibody species specificity on dualcolor STORM imaging. In the classical erythrocyte skeleton model, based on the mutually exclusive localization between the Nterminal and Cterminal of βspectrin, we performed twocolor STORM imaging of them by labeling them with lowand highadsorption secondary antibodies, respectively. A comparison with crosscorrelation data from simulations revealed that lowadsorption secondary antibodies led to colocalization artifacts in

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Multicolor Single-Molecule Localization Super-Resolution Microscopy

Cited by 2 publications

References 0 publications

Multicolor single-molecule localization microscopy: review and prospect

Multicolor single-molecule localization microscopy: review and prospect

二抗物种特异性对双色storm成像的影响（特邀）

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