Gradual labeling with fluorogenic probes for MINFLUX nanoscopy of densely packed structures
yao, l.; Si, D.; Chen, L.; Li, S.; Guan, J.; Ma, J.; zhang, q.; wang, j.; wang, l.; Gu, M.
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Single-molecule localization in densely packed structures represents a formidable challenge in nanoscopy. Here we develop a gradual labeling method with fluorogenic probes for MINFLUX nanoscopy (GLF-MINFLUX), with which individual proteins in packed structures can be sequential illuminated and quantitatively localized at the nanoscale solely by adjusting probe concentration. With a 1.7-fold localization precision improvement and 2.2-fold faster acquisition, GLF-MINFLUX enables dual-channel nanoscale imaging of densely neuronal microtubules and microfilaments, and three-dimensional quantitative mapping of individual TOM20 proteins in mitochondrial clusters.
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