Near-infrared STED nanoscopy with an engineered bacterial phytochrome

Kamper, Maria; Ta, Haisen; Jensen, Nickels; Hell, Stefan W.; Jakobs, Stefan

doi:10.1038/s41467-018-07246-2

Cited by 40 publications

(57 citation statements)

References 33 publications

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“…The brightness of the optimized emiRFPs allowed to perform NIR STED with the widely used 640 nm excitation laser and without addition of exogenous BV chromophore, in contrast to STED performed earlier with SNIFP protein with low quantum yield (2.2%) of DrBphP origin 33 . The NIR FPs are excited at or close to their peak by 640-670 nm lasers.…”

Section: Articlementioning

confidence: 99%

A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales

et al. 2020

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Bright monomeric near-infrared (NIR) fluorescent proteins (FPs) are in high demand as protein tags for multicolor microscopy and in vivo imaging. Here we apply rational design to engineer a complete set of monomeric NIR FPs, which are the brightest genetically encoded NIR probes. We demonstrate that the enhanced miRFP series of NIR FPs, which combine high effective brightness in mammalian cells and monomeric state, perform well in both nanometer-scale imaging with diffraction unlimited stimulated emission depletion (STED) microscopy and centimeter-scale imaging in mice. In STED we achieve~40 nm resolution in live cells. In living mice we detect~10 5 fluorescent cells in deep tissues. Using spectrally distinct monomeric NIR FP variants, we perform two-color live-cell STED microscopy and two-color imaging in vivo. Having emission peaks from 670 nm to 720 nm, the next generation of miRFPs should become versatile NIR probes for multiplexed imaging across spatial scales in different modalities.

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Section: Articlementioning

confidence: 99%

A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales

et al. 2020

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“…The OAM of light has led to a myriad of new physical effects and various applications particularly at the microscopic scale. This includes optical trapping and manipulation, optical telecommunications, quantum physics, and “super‐resolution” microscopy with a spatial resolution beyond the diffraction limit . New applications for OAM fields have also been proposed in environmental optics and free‐space telecommunication.…”

Section: Introductionmentioning

confidence: 99%

A New Twist for Materials Science: The Formation of Chiral Structures Using the Angular Momentum of Light

Omatsu

Miyamoto

Toyoda

et al. 2019

Advanced Optical Materials

119

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and various applications particularly at the microscopic scale. This includes optical trapping and manipulation, [4][5][6] optical telecommunications, [7,8] quantum physics, [9] and "super-resolution" microscopy with a spatial resolution beyond the diffraction limit. [10][11][12] New applications for OAM fields have also been proposed in environmental optics and free-space telecommunication. As an example OAM states can potentially propagate though air turbulence with lower degradation than a conventional Gaussian beam. [13,14] New physical effects include studies such as the rotational Doppler effect originating from interactions of such fields and rotating objects that may provide new technologies for remote sensing of rotating bodies in astrophysics. [15,16] In this area of original physical demonstrations, recent studies have shown that irradiation by such an OAM field can twist materials, such as metal, silicon, azo-polymer, and even a liquid-phase resin with the help of spin angular momentum (SAM) of circularly polarized light with the rotating electric field. This can thus shape helical nano/microstructures. [17][18][19][20][21][22][23][24] Going beyond fundamental aspects, twisted materials created by such OAM fields may provide a new understanding of interactions between optical fields and matter on the subwavelength scale that may reveal new physics, e.g., spin-orbit coupling effects or unconventional optomechanical effects for moving beyond traditional forms of optical manipulation. Furthermore, twisted materials created by such OAM fields, will provide a new understanding of interactions between optical fields and matter on a subwavelength scale. For example, the twisted materials manifest the role of both SAM and OAM of light fields and the coupling of SAM and OAM (so-called spin-orbit coupling) effects. This coupling is key not only for understanding fundamental physics but also for controlling optical material structures.Conventional optical tweezers rely on the field gradients near the diffraction limited focus of a laser beam to hold mesoscopic particles solely with focused light beams. However, the technique often fails to efficiently trap and manipulate particles at the nanoscale because the gradient force scales with the volume of the particle (for a dielectric object) and is proportional to the particle's polarizability. In this domain, advanced materials science and nanofabrication technologies have made remarkable progress in structured devices, e.g., photonic crystals, metamaterials, and metasurfaces. These devices offer novel trapping geometries to enhance the interaction between optical fields and materials at the nanoscale. Furthermore, these devices allow the control of Recent work has shown that irradiation with light possessing orbital angular momentum (OAM) and an associated phase singularity, that is an optical vortex, twists a variety of materials. These include silicon, azo-polymer, and even liquid-phase resins to form various helically structured materials. This article provides...

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“…On the down side, STED depletion beam deploys some of the highest laser power among all SRM techniques, raising concerns over phototoxicity to biological samples and unrecoverable photodestruction of FPs (phototoxicity and FP photodestruction are often loosely quoted under the collective term of photodamage). Despite that, time-lapse STED imaging in living cells has been demonstrated on several occasions [24,25]. For functional SRM, a H 2 O 2 indicator based on cpYFP, named HyPer2, has been successfully observed with STED [26].…”

Section: Sense and Sensibility: Anatomy Of Genetically Encoded Indmentioning

confidence: 99%

“…Recently, effective brightness and monomeric property of near-infrared biliproteins have been greatly improved to accommodate bioimaging applications in mammalian cells [29,30,31]. For SRM, a photostable biliprotein with far-red emission, named SNIFP, has been successfully applied to the imaging of cytoskeleton and nucleus pores in living cells (notably, low quantum yield of SNIFP is likely offset by the combination of strong laser excitation in STED and sensitive detector) [25]. Compared to STED, SIM requires less aggressive illumination scheme, and is arguably more amiable towards living samples [32].…”

Section: Sense and Sensibility: Anatomy Of Genetically Encoded Indmentioning

confidence: 99%

Fluorescent Protein-Based Indicators for Functional Super-Resolution Imaging of Biomolecular Activities in Living Cells

Matsuda

et al. 2019

IJMS

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Super-resolution light microscopy (SRM) offers a unique opportunity for diffraction-unlimited imaging of biomolecular activities in living cells. To realize such potential, genetically encoded indicators were developed recently from fluorescent proteins (FPs) that exhibit phototransformation behaviors including photoactivation, photoconversion, and photoswitching, etc. Super-resolution observations of biomolecule interactions and biochemical activities have been demonstrated by exploiting the principles of bimolecular fluorescence complementation (BiFC), points accumulation for imaging nanoscale topography (PAINT), and fluorescence fluctuation increase by contact (FLINC), etc. To improve functional nanoscopy with the technology of genetically encoded indicators, it is essential to fully decipher the photo-induced chemistry of FPs and opt for innovative indicator designs that utilize not only fluorescence intensity but also multi-parametric readouts such as phototransformation kinetics. In parallel, technical improvements to both the microscopy optics and image analysis pipeline are promising avenues to increase the sensitivity and versatility of functional SRM.

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Near-infrared STED nanoscopy with an engineered bacterial phytochrome

Cited by 40 publications

References 33 publications

A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales

A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales

A New Twist for Materials Science: The Formation of Chiral Structures Using the Angular Momentum of Light

Fluorescent Protein-Based Indicators for Functional Super-Resolution Imaging of Biomolecular Activities in Living Cells

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