Enzymatic activation of nitro-aryl fluorogens in live bacterial cells for enzymatic turnover-activated localization microscopy

Lee, Marissa K.; Williams, Jarrod; Twieg, Robert J.; Rao, Jianghong; Moerner, W. E.

doi:10.1039/c2sc21074f

Cited by 58 publications

(58 citation statements)

References 45 publications

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“…7. There are photochemical methods for single-fluorophore turn-on 109 and even enzymatic methods for turn-on which may be controlled by the concentration of substrate and the enzymatic rate 110 . The experimenter must actively choose some method to control the emitting concentration.…”

Section: Super-resolution Imaging Based On Single Moleculesmentioning

confidence: 99%

Single-molecule spectroscopy and imaging over the decades

2015

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As of 2015, it has been 26 years since the first optical detection and spectroscopy of single molecules in condensed matter. This area of science has expanded far beyond the early low temperature studies in crystals to include single molecules in cells, polymers, and in solution. The early steps relied upon high-resolution spectroscopy of inhomogeneously broadened optical absorption profiles of molecular impurities in solids at low temperatures. Spectral fine structure arising directly from the position-dependent fluctuations of the number of molecules in resonance led to the attainment of the single-molecule limit in 1989 using frequency-modulation laser spectroscopy. In the early 1990's, a variety of fascinating physical effects were observed for individual molecules, including imaging of the light from single molecules as well as observations of spectral diffusion, optical switching and the ability to select different single molecules in the same focal volume simply by tuning the pumping laser frequency. In the room temperature regime, researchers showed that bursts of light from single molecules could be detected in solution, leading to imaging and microscopy by a variety of methods. Studies of single copies of the green fluorescent protein also uncovered surprises, especially the blinking and photoinduced recovery of emitters, which stimulated further development of photoswitchable fluorescent protein labels. All of these early steps provided important fundamentals underpinning the development of super-resolution microscopy based on single-molecule localization and active control of emitting concentration. Current thrust areas include extensions to three-dimensional imaging with high precision, orientational analysis of single molecules, and direct measurements of photodynamics and transport properties for single molecules trapped in solution by suppression of Brownian motion. Without question, a huge variety of studies of single molecules performed by many talented scientists all over the world have extended our knowledge of the nanoscale and microscopic mechanisms previously hidden by ensemble averaging.

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Section: Super-resolution Imaging Based On Single Moleculesmentioning

confidence: 99%

Single-molecule spectroscopy and imaging over the decades

2015

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“…[53][54][55][56][57] To obtain the colorimetric and long-wavelength fluorophore, the hydroxyl moiety (oxygen anions in physiological conditions) with strong electron-donor ability has been introduced. Additionally, it is reported that arylboronate can react with OClto yield hydroxyl derivatives much faster than does H 2 O 2 .…”

Section: Introductionmentioning

confidence: 99%

A novel visual and far-red fluorescent dual-channel probe for the rapid and sensitive detection of hypochlorite in aqueous solution and living cells

Shu

Yan

Wang

et al. 2015

Sensors and Actuators B: Chemical

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“…In this way, the probe (nitro-aryl fluorogen 8 , Fig. 5c,d) acts as a readout of enzyme activity in live cells, enabling visualization of the location of nitroreductase in an approach called enzymatic-turnover-activated localization microscopy (ETALM) 93 .…”

Section: Additional Small-molecule Probesmentioning

confidence: 99%

Progress and prospects for small-molecule probes of bacterial imaging

Kocaoglu

Carlson

2016

Nat Chem Biol

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Fluorescence microscopy is an essential tool for the exploration of cell growth, division, transcription and translation in eukaryotes and prokaryotes alike. Despite the rapid development of techniques to study bacteria, the size of these organisms (1–10 μm) and their robust and largely impenetrable cell envelope present major challenges in imaging experiments. Fusion-based strategies, such as attachment of the protein of interest to a fluorescent protein or epitope tag, are by far the most common means for examining protein localization and expression in prokaryotes. While valuable, the use of genetically encoded tags can result in mislocalization or altered activity of the desired protein, does not provide a readout of the catalytic state of enzymes and cannot enable visualization of many other important cellular components, such as peptidoglycan, lipids, nucleic acids or glycans. Here, we highlight the use of biomolecule-specific small-molecule probes for imaging in bacteria.

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Enzymatic activation of nitro-aryl fluorogens in live bacterial cells for enzymatic turnover-activated localization microscopy

Cited by 58 publications

References 45 publications

Single-molecule spectroscopy and imaging over the decades

Single-molecule spectroscopy and imaging over the decades

A novel visual and far-red fluorescent dual-channel probe for the rapid and sensitive detection of hypochlorite in aqueous solution and living cells

Progress and prospects for small-molecule probes of bacterial imaging

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