A5035896249- Advanced Fluorescence Microscopy Techniques
- Advanced Electron Microscopy Techniques and Applications
- Cell Image Analysis Techniques
- Cancer Research and Treatments
- Photoacoustic and Ultrasonic Imaging
- DNA Repair Mechanisms
- DNA and Nucleic Acid Chemistry
- Molecular Biology Techniques and Applications
- Bacterial Genetics and Biotechnology
- Near-Field Optical Microscopy
- ATP Synthase and ATPases Research
- Nanoplatforms for cancer theranostics
- Photoreceptor and optogenetics research
- Single-cell and spatial transcriptomics
- Diffusion and Search Dynamics
- Advanced Biosensing Techniques and Applications
University of Edinburgh
2022-2025
Edinburgh College
2024
Institut de Biologie Structurale
2017-2023
Université Grenoble Alpes
2018-2022
CEA Grenoble
2022
Commissariat à l'Énergie Atomique et aux Énergies Alternatives
2022
Centre National de la Recherche Scientifique
2018-2022
Green-to-red photoconvertible fluorescent proteins (PCFPs) are key players in advanced microscopy schemes such as photoactivated localization (PALM). Whereas photoconversion and red-state blinking PCFPs have been studied intensively, their green-state photophysical behavior has received less attention. Yet dark states green can become strongly populated PALM exert an indirect but considerable influence on the quality of data recorded red channel. Furthermore, photoswitching be used directly...
Repairing DNA damage is of primary importance for all living organisms. double-strand breaks (DSBs) are one the most serious types damage, as they lead to loss genetic information and death when not repaired. In E. coli , recognised processed by RecBCD complex, which initiates repair homologous recombination. Although dynamics downstream has been well characterised, it still unclear how long this complex stays attached what triggers its dissociation in vivo . To answer these questions, we...
Single-molecule localization microscopy (SMLM) at cryogenic temperature opens new avenues to investigate intact biological samples the nanoscale and perform cryo-correlative studies. Genetically encoded fluorescent proteins (FPs) are markers of choice for cryo-SMLM, but their reduced conformational flexibility below glass-transition hampers efficient cryo-photoswitching. We investigated cryo-switching rsEGFP2, one most reversibly switchable ambient due facile cis–trans isomerization...
Green-to-red photoconvertible fluorescent proteins (PCFPs) are widely employed as markers in photoactivated localization microscopy (PALM). However, their highly complex photophysical behavior complicates usage. The fact that only a limited fraction of PCFP ensemble can form the photoconverted state upon near-UV light illumination, termed photoconversion efficiency (PCE), lowers achievable spatial resolution PALM and creates undercounting errors quantitative counting applications. Here, we...
Green-to-red photoconvertible fluorescent proteins (PCFPs) such as mEos2 and its derivatives are widely used in PhotoActivated Localization Microscopy (PALM). However, the complex photophysics of these genetically encoded markers complicates quantitative analysis PALM data. Here, we show that intense 561 nm light (∼1 kW/cm2) typically to localize single red molecules considerably affects green-state by populating at least two reversible dark states. These states retard green-to-red...
Bioimage analysis (BIA), a crucial discipline in biological research, overcomes the limitations of subjective microscopy through creation and application quantitative reproducible methods. The establishment dedicated BIA support within academic institutions is vital to improving research quality efficiency can significantly advance scientific discovery. However, lack training resources, limited career paths insufficient recognition contributions made by bioimage analysts prevent full...
Abstract Efficient DNA repair is crucial for maintaining genome integrity and ensuring cell survival. In Escherichia coli , RecBCD plays a role in processing ends following double-strand break (DSB) to initiate by homologous recombination. While has been extensively studied vitro less known about how it contributes rapid efficient living bacteria. Here, we perform single-molecule microscopy investigate real-time E. . We quantify RecB mobility monitor the induction of damage response (SOS...
Green-to-red photoconvertible fluorescent proteins repeatedly enter dark states, causing interrupted tracks in single-particle-tracking localization microscopy (sptPALM). We identified a long-lived state photoconverted mEos4b that results from isomerization of the chromophore and efficiently absorbs cyan light. Addition weak 488-nm light swiftly reverts this to state. This strategy largely eliminates slow blinking enables recording significantly longer sptPALM with minimum effort.
ABSTRACT Single-molecule-localization-microscopy (SMLM) at cryogenic temperature opens new avenues to investigate intact biological samples the nanoscale and perform cryo-correlative studies. Genetically encoded fluorescent proteins (FPs) are markers of choice for cryo-SMLM, but their reduced conformational flexibility below glass transition hampers efficient photoswitching low temperature. We investigated cryo-switching rsEGFP2, one most reversibly switchable protein ambient due facile...
Green-to-red photoconvertible fluorescent proteins (PCFPs) are widely employed as markers in photoactivated localization microscopy (PALM). However, their highly complex photophysical behavior complicates usage. The fact that only a limited fraction of PCFP ensemble can form the photoconverted state upon near-UV light illumination, termed photoconversion efficiency (PCE), lowers achievable spatial resolution PALM and creates undercounting errors quantitative counting applications. Here, we...