Super-resolution fluorescence microscopy for investigating bacterial macromolecular complexes DOI Open Access
Alexander Carsten, Manuel Wolters, Martin Aepfelbacher

et al.

Authorea (Authorea), Journal Year: 2023, Volume and Issue: unknown

Published: Aug. 5, 2023

Super-resolution fluorescence microscopy techniques developed over the past two decades have pushed resolution limit for fluorescently labeled molecules into nanometer range. These potential to study bacterial macromolecular complexes such as secretion systems with single-molecule on a millisecond time scale. Here we review recent applications of super-resolution in molecular bacteriology focus systems. We also describe MINFLUX nanoscopy, relatively new technique that promises one day produce movies machines action.

Language: Английский

Imaging-free, few-shot, three-dimensional focusing on point-like emitters in confocal microscopy DOI

Swetapadma Sahoo,

Junyue Jiang,

Jaden Li

et al.

Published: Jan. 1, 2024

We introduce a rapid, noise-robust, three-dimensional focusing framework for as-is confocal microscopes. show automated real-time on nanoscale emitters SNR down to 1, and position tracking with precision below 10 nm.

Language: Английский

Citations

0
Czy można obejść prawa fizyki? O metodach super-rozdzielczej mikroskopii fluorescencyjnej DOI Open Access
Adrian Rüfli

Postępy Biochemii, Journal Year: 2024, Volume and Issue: 70(2), P. 139 - 149

Published: July 1, 2024

Biological sciences are increasingly uncovering the foundations of life in greater detail, made possible by development research methods enabling exploration at nanometer scale. Optical microscopy, a field with significant contribution to current knowledge, is inherently limited Abbe limit, stemming from fundamental wave properties light. Through efforts scientists, this limit can be circumvented, as evidenced STED and MINFLUX techniques. allows imaging resolution down 40 nm, while enables fine 2 nm. Both techniques require labelling biological molecular targets fluorescent markers enable living cells, facilitating study dynamic processes. This article provides an introduction super-resolution MINFLUX, demonstrating their utility through example studying kinesin movement along microtubules using technique.

Language: Английский

Citations

0
Red-Emitting Fluorophores with Tailored Properties for Microscopy and Nanoscopy DOI Creative Commons
Kirill Kolmakov, Massimiliano Lucidi, Alexey V. Nizovtsev

et al.

IntechOpen eBooks, Journal Year: 2024, Volume and Issue: unknown

Published: June 20, 2024

Extended series of red-emitting rhodamine dyes were synthesized and tested. This revealed the real factors determining quality STED (stimulated emission depletion) imaging, dye photostability in general, stability corresponding reactive labels. Previously overlooked underestimated observations are now being highlighted discussed. So some very important syntheses-related aspects. The studies also expanded on far-red emitting oxazine silicon dyes. An intriguing discovery emerged when certain fluorophores, which originally designed for STED, demonstrated an ability to discern between viable dead bacterial cells. Recent publications by Lucidi co-workers showed that positively charged allow imaging membranes using both conventional techniques, example, confocal laser scanning microscopy (CLSM), STED. opens avenues investigating membrane biogenesis diagnostics.

Language: Английский

Citations

0
Bioorthogonal caging-group-free photoactivatable probes for minimal-linkage-error nanoscopy DOI Creative Commons
Ayse Aktalay, Richard Lincoln,

Lukas Heynck

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: June 22, 2023

ABSTRACT Here we describe highly compact, click compatible, and photoactivatable dyes for super-resolution fluorescence microscopy (nanoscopy). By combining the xanthone (PaX) core with a tetrazine group, achieve minimally sized sensitive molecular dyads selective labeling of unnatural amino acids introduced by genetic code expansion. We exploit excited state quenching properties group to attenuate photoactivation rates PaX, further reduce overall emission photogenerated fluorophore, providing two mechanisms selectivity off-target signal. Coupled MINFLUX nanoscopy, demonstrate our in minimal-linkage-error imaging vimentin filaments, demonstrating scale precision fluorophore positioning.

Language: Английский

Citations

0
Super-resolution fluorescence microscopy for investigating bacterial macromolecular complexes DOI Open Access
Alexander Carsten, Manuel Wolters, Martin Aepfelbacher

et al.

Authorea (Authorea), Journal Year: 2023, Volume and Issue: unknown

Published: Aug. 5, 2023

Super-resolution fluorescence microscopy techniques developed over the past two decades have pushed resolution limit for fluorescently labeled molecules into nanometer range. These potential to study bacterial macromolecular complexes such as secretion systems with single-molecule on a millisecond time scale. Here we review recent applications of super-resolution in molecular bacteriology focus systems. We also describe MINFLUX nanoscopy, relatively new technique that promises one day produce movies machines action.

Language: Английский

Citations

0