Wide-field fluorescence microscopy articles within Nature Communications

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  • Article
    | Open Access

    Implementing point spread function (PSF) engineering in high-throughput microscopy has proved challenging. Here, the authors propose a compact PSF engineering approach, which allows for enhanced depth of field and for the recovery of 3D information using single snapshots.

    • Nadav Opatovski
    • , Elias Nehme
    •  & Yoav Shechtman

  • Article
    | Open Access

    The neurophysiological mechanisms of deep brain stimulation remain poorly understood. Through fluorescence voltage imaging of individual hippocampal neurons in awake mice, the authors show that deep brain stimulation causes membrane depolarization that impairs a neuron’s ability to respond to intrinsic network activity patterns and optogenetic somatic depolarization, thereby creating an informational lesion.

    • Eric Lowet
    • , Krishnakanth Kondabolu
    •  & Xue Han

  • Article
    | Open Access

    Chromatin compaction affects many nuclear processes yet compaction levels at individual genomic loci have been notoriously difficult to assess. Here, Ana Mota and co-authors from the Bienko-Crosetto Lab present FRET-FISH for probing chromatin compaction at selected loci in single cells.

    • Ana Mota
    • , Szymon Berezicki
    •  & Magda Bienko

  • Article
    | Open Access

    Here, the authors develop a UV-compatible photonic integrated circuit for structured illumination microscopy on a conventional wide-field microscope. Operating at a wavelength of 360 nm, they generate switchable far-field fringe patterns, and demonstrate autofluorescence imaging of yeast cells.

    • Chupao Lin
    • , Juan Santo Domingo Peñaranda
    •  & Nicolas Le Thomas

  • Article
    | Open Access

    Biosensors often report relative rather than absolute values. Here the authors report a method that utilises the photochromic properties of biosensors to provide an absolute measure of the analyte concentration or activity: photochromism-enabled absolute quantification (PEAQ) biosensing.

    • Franziska Bierbuesse
    • , Anaïs C. Bourges
    •  & Peter Dedecker

  • Article
    | Open Access

    The authors demonstrate accurate localization in three dimensions by comprehensive calibration of an ordinary microscope, exploiting the latent information of intrinsic aberrations. Rigid transformation of the emitter positions tests the method and enables measurements in six degrees of freedom.

    • Craig R. Copeland
    • , Craig D. McGray
    •  & Samuel M. Stavis

  • Article
    | Open Access

    Uniform illumination is a prerequisite for quantitative analyses in both classical fluorescence microscopy and single molecule localisation microscopy. Here, the authors introduce ASTER, an illumination technique that generates uniform illumination over large and adaptable fields of view, compatible with epifluorescence, HiLo and TIRF illumination schemes.

    • Adrien Mau
    • , Karoline Friedl
    •  & Sandrine Lévêque-Fort

  • Article
    | Open Access

    Performing multiple FRET measurements at once can be challenging. Here the authors report a method to discriminate between overlapping FRET pairs, even if the fluorophores display almost identical absorption and emission spectra, based on the photochromism of the donor fluorophores.

    • Thijs Roebroek
    • , Wim Vandenberg
    •  & Peter Dedecker

  • Article
    | Open Access

    Structured illumination microscopy is usually limited to 2 times spatial resolution improvement over the diffraction limit. Here, the authors introduce a metamaterial structure to generate speckle-like sub-diffraction limit illumination patterns in the near field, and achieve a 7-fold resolution improvement down to 40 nm.

    • Yeon Ui Lee
    • , Junxiang Zhao
    •  & Zhaowei Liu

  • Article
    | Open Access

    Fluorescent protein reporters based on GFP exist, but have intrinsic disadvantages. Here the authors incorporate pH, Ca2+ and protein–protein interaction sensing modalities into de novo designed mini-fluorescence-activating proteins (mFAPs), with increased photostability and smaller size, which bind a range of DFHBI chromophore variants.

    • Jason C. Klima
    • , Lindsey A. Doyle
    •  & David Baker

  • Article
    | Open Access

    Here, the authors generalize cumulant analysis by extending it into the spectral domain to allow multicolour super-resolution optical fluctuation imaging. The simultaneous acquisition of two spectral channels followed by spectral cross-cumulant analysis and unmixing allows denser spectral and spatial sampling of the super-resolved image.

    • K. S. Grußmayer
    • , S. Geissbuehler
    •  & T. Lasser

  • Article
    | Open Access

    Performing homo-FRET measurements in cells using a fluorescence microscope is challenging, especially when using high numerical aperture objective lenses. Here the authors present a method for improved homo-FRET measurements based on anisotropy changes in photoswitchable fluorescent proteins.

    • Namrata Ojha
    • , Kristin H. Rainey
    •  & George H. Patterson

  • Article
    | Open Access

    Nanosecond imaging techniques, such as fluorescence lifetime imaging microscopy (FLIM), are limited by low efficiency of current detectors. Here, the authors implement an electro-optic approach using Pockels cells for wide-field image gating and demonstrate high throughput FLIM on standard camera sensors.

    • Adam J. Bowman
    • , Brannon B. Klopfer
    •  & Mark A. Kasevich

  • Article
    | Open Access

    Time-lapse imaging studies of more than a day in the fly brain have been infeasible until now. Here the authors present a laser microsurgery approach to create a permanent window in the fly cuticle to enable time-lapse imaging of neural architecture and dynamics for up to 10–50 days.

    • Cheng Huang
    • , Jessica R. Maxey
    •  & Mark J. Schnitzer

  • Article
    | Open Access

    Imaging of inorganic dispersions in organic-inorganic composites is typically performed using electron microscopy. Here, the authors show that surfactants with attached aggregation-induced emission fluorophores allow simple fluorescence imaging of the spatial distribution of the inorganic filler dispersion.

    • Weijiang Guan
    • , Si Wang
    •  & Ben Zhong Tang

  • Article
    | Open Access

    Increasing the resolution of fluorescence microscopy is a fundamental need for modern cell biology. Lanzanò et al.demonstrate that arbitrary spatial resolution is, in principle, possible by encoding the fluorophore's spatial distribution information in the temporal dynamics of the fluorophore's transition.

    • Luca Lanzanò
    • , Iván Coto Hernández
    •  & Giuseppe Vicidomini

  • Article
    | Open Access

    Super-resolution optical fluctuation imaging provides 3D images of biological specimens via blinking fluorophores. Geissbuehler et al. present a multiplexed version of this method that captures images at multiple focal planes simultaneously, reducing the acquisition time compared with standard approaches.

    • Stefan Geissbuehler
    • , Azat Sharipov
    •  & Marcel Leutenegger

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