Carboxylated Photoswitchable Diarylethenes for Biolabeling and Super‐Resolution RESOLFT Microscopy

Roubinet, Benoît; Bossi, Mariano L.; Alt, Philipp; Leutenegger, Marcel; Shojaei, Heydar; Schnorrenberg, Sebastian; Nizamov, Shamil; Irie, Masahiro; Belov, Vladimir N.; Hell, Stefan W.

doi:10.1002/anie.201607940

Cited by 135 publications

(136 citation statements)

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“…Compound 1b presents Φ fluo /Φ on→off < 300; with a photon collection efficiency (CEF) of ∼ 10%, and around 30 photons per burst (i.e. 29 For this reason, this compound is not suitable for STORM; and slowing the off-switching rate by at least one order of magnitude lower Φ on→off is necessary.…”

Section: Resultsmentioning

confidence: 99%

“…37 The switching photochromic unit is intrinsically not water-soluble, but the new DAE derivatives allowed a viable and promising approach to be realized. 29 Images of immunolabeled cells, with a spatial resolution beyond the diffraction barrier, were obtained using conjugates of compounds 1a,b in RESOLFT microscopy (reversible saturable [switchable] optically linear fluorescence transitions), a coordinate-targeted imaging technique. They exhibited fluorescence modulation in aqueous buffers at biologically relevant pH values (as free markers and conjugates with secondary antibodies).…”

Section: Introductionmentioning

confidence: 99%

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Fluorescent Photoswitchable Diarylethenes for Biolabeling and Single-Molecule Localization Microscopies with Optical Superresolution

et al. 2017

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A modular assembly of water-soluble diarylethenes (DAEs), applicable as biomarkers for optical nanoscopy, is reported. Reversibly photoswitchable 1,2-bis(2-alkyl-6-phenyl-1-benzothiophene-1,1-dioxide-3-yl)perfluorocyclopentenes possessing a fluorescent "closed" form were decorated with one or two methoxy group(s) attached to the para-position(s) of phenyl ring(s) and two, four, or eight carboxylic acid groups. Antibody conjugates of these DAEs feature low aggregation, efficient photoswitching in aqueous buffers, specific staining of cellular structures, and photophysical properties (high emission efficiencies and low cycloreversion quantum yields) enabling their application in superresolution microscopy. Images of tubulin, vimentin, and nuclear pore complexes are presented. The superresolution images can also be acquired by using solely 488 nm light without additional photoactivation with UV light. These DAEs exhibit reversible photoswitching without requiring any additives to the imaging media and open new paths toward the modular design of fluorescent dyes for bioimaging with optical superresolution.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fluorescent Photoswitchable Diarylethenes for Biolabeling and Single-Molecule Localization Microscopies with Optical Superresolution

et al. 2017

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“…A fundamental concept, MINFIELD will also be applicable to other types of coordinate-targeted superresolution variants, such as ground-state depletion (26) and reversible saturable/switchable optical fluorescence transitions (RESOLFT) nanoscopy using photoswitchable fluorescent proteins (27)(28)(29) and switchable (photochromic) synthetic-organic fluorophores (30). Moreover, MINFIELD will be relevant for superresolution modalities that use light patterns for switching fluorophores on rather than off.…”

Section: Resultsmentioning

confidence: 99%

Strong signal increase in STED fluorescence microscopy by imaging regions of subdiffraction extent

Göttfert

Pleiner

Heine

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Photobleaching remains a limiting factor in superresolution fluorescence microscopy. This is particularly true for stimulated emission depletion (STED) and reversible saturable/switchable optical fluorescence transitions (RESOLFT) microscopy, where adjacent fluorescent molecules are distinguished by sequentially turning them off (or on) using a pattern of light formed as a doughnut or a standing wave. In sample regions where the pattern intensity reaches or exceeds a certain threshold, the molecules are essentially off (or on), whereas in areas where the intensity is lower, that is, around the intensity minima, the molecules remain in the initial state. Unfortunately, the creation of on/off state differences on subdiffraction scales requires the maxima of the intensity pattern to exceed the threshold intensity by a large factor that scales with the resolution. Hence, when recording an image by scanning the pattern across the sample, each molecule in the sample is repeatedly exposed to the maxima, which exacerbates bleaching. Here, we introduce MINFIELD, a strategy for fundamentally reducing bleaching in STED/RESOLFT nanoscopy through restricting the scanning to subdiffraction-sized regions. By safeguarding the molecules from the intensity of the maxima and exposing them only to the lower intensities (around the minima) needed for the off-switching (on-switching), MINFIELD largely avoids detrimental transitions to higher molecular states. A bleaching reduction by up to 100-fold is demonstrated. Recording nanobody-labeled nuclear pore complexes in Xenopus laevis cells showed that MINFIELD-STED microscopy resolved details separated by <25 nm where conventional scanning failed to acquire sufficient signal.fluorescence nanoscopy | STED microscopy | photobleaching | superresolution

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“…As a result, the development of water-compatible photoswitchable logic gates is still a challenging task. [22][23][24][25] Recently, we have demonstrated that the glycosylation of hydrophobic organic dyes is a promising strategy to overcome the water insolubility of traditional organic dyes, achieving full-aqueous sensing and targeted cell imaging. 26 Here we show that the introduction of a carbohydrate group to diarylethenes can lead to the development of a water-compatible reversible photochromic system with logicgate operation.…”

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confidence: 99%

Photochromism and molecular logic gate operation of a water-compatible bis-glycosyl diarylethene

Chai

James

et al. 2017

Chem. Commun.

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Carboxylated Photoswitchable Diarylethenes for Biolabeling and Super‐Resolution RESOLFT Microscopy

Cited by 135 publications

References 37 publications

Fluorescent Photoswitchable Diarylethenes for Biolabeling and Single-Molecule Localization Microscopies with Optical Superresolution

Fluorescent Photoswitchable Diarylethenes for Biolabeling and Single-Molecule Localization Microscopies with Optical Superresolution

Strong signal increase in STED fluorescence microscopy by imaging regions of subdiffraction extent

Photochromism and molecular logic gate operation of a water-compatible bis-glycosyl diarylethene

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