Evidence map›Paper›PMID 40397744›Full record

ArticleScience advances2025

Gradual labeling with fluorogenic probes: A general method for MINFLUX imaging and tracking.

Longfang Yao, Dongjuan Si, Liwen Chen, Shu Li, Jiaxin Guan, Qiming Zhang, Jing Wang, Jiong Ma, Lu Wang, Min Gu

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Science advances, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors.

Longfang YaoSchool of Artificial Intelligence Science and Technology, University of Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, China.ORCID 0000-0002-1653-5726
Dongjuan SiMOE Key Laboratory of Smart Drug Delivery, MOE Innovative Center for New Drug Development of Immune Inflammatory Diseases, School of Pharmacy, Fudan University, Shanghai 200433, China.
Liwen ChenDepartment of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-precision Optical Manufacturing, Fudan University, 220 Handan Road, Shanghai 200433, China.ORCID 0000-0002-7271-2467
Shu LiSchool of Artificial Intelligence Science and Technology, University of Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, China.ORCID 0009-0009-6845-8935
Jiaxin GuanSchool of Artificial Intelligence Science and Technology, University of Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, China.
Qiming ZhangSchool of Artificial Intelligence Science and Technology, University of Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, China.ORCID 0000-0003-0067-1210
Jing WangSchool of Artificial Intelligence Science and Technology, University of Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, China.
Jiong MaDepartment of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-precision Optical Manufacturing, Fudan University, 220 Handan Road, Shanghai 200433, China.ORCID 0000-0003-3632-4624
Lu WangMOE Key Laboratory of Smart Drug Delivery, MOE Innovative Center for New Drug Development of Immune Inflammatory Diseases, School of Pharmacy, Fudan University, Shanghai 200433, China.ORCID 0000-0001-8412-2985
Min GuSchool of Artificial Intelligence Science and Technology, University of Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, China.ORCID 0000-0003-4078-253X

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Minimal photon fluxes (MINFLUX) nanoscopy excels in nanoscale protein studies but lacks a universal method for simultaneous imaging and live-cell tracking in dense cellular environments. Here, we developed a general strategy, gradual labeling with fluorogenic probes for MINFLUX (GLF-MINFLUX) imaging and tracking. In GLF-MINFLUX, membrane-permeable small-molecule fluorogenic dye with protein-induced "off/on" switching is gradually labeled, located, and bleached, enabling sequential positioning and tracking of individual proteins. GLF-MINFLUX reveals continuous microtubules with 2.6-nanometer localization precision, offering substantially improved precision (1.7-fold), acquisition (2.2-fold), and target density (3-fold) compared to conventional MINFLUX with Alexa Fluor 647. GLF-MINFLUX also enabled the three-dimensional localization of translocase of the outer mitochondrial membrane 20 proteins within mitochondrial clusters and dual-channel nanoscale imaging of endogenous neuronal microtubules and microfilaments. GLF-MINFLUX allowed live-cell single-protein tracking with 7.8-nanometer precision at ~200-microsecond temporal resolution, revealing distinct diffusion behaviors and rates between the basal membrane and filopodia. GLF-MINFLUX, requiring only tuning of probe concentration, offers molecular-level insights into protein functions.

Indexed as

Cell TrackingFluorescent DyesStaining and LabelingAnimalsHumansMicroscopy, FluorescenceMicrotubulesPhotonsFluorescent Dyes

Identifiers

PMID40397744
PMCPMC12094232

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.