Microscopy articles within Nature Methods

Featured

  • Brief Communication |

    Stimulated Raman scattering (SRS) microscopy is a quantitative, label-free imaging method to map fat distribution and accumulation with high spatial resolution and sensitivity at both cellular and organism levels.

    • Meng C Wang
    • , Wei Min
    •  & X Sunney Xie

  • Brief Communication |

    In vivo calcium imaging at multiple depths simultaneously is shown using multifocal two-photon microscopy and spatiotemporal multiplexing. This technique involves scanning the sample with multiple beams in parallel at different axial planes and is applied to monitor neuronal network activity in multiple cortical layers of an anesthetized mouse.

    • Adrian Cheng
    • , J Tiago Gonçalves
    •  & Carlos Portera-Cailliau

  • News & Views |

    Stabilization with a suction window permits in vivo imaging of the mouse lung vasculature with video-rate two-photon microscopy.

    • Jahar Bhattacharya

  • Article |

    Fast, two-photon intravital imaging of a mechanically stabilized and physiologically intact preparation of the mouse lung is reported. It is used to monitor immune cells in the lung under normal and injured conditions.

    • Mark R Looney
    • , Emily E Thornton
    •  & Matthew F Krummel

  • Brief Communication |

    A soft X-ray microscope design using partially incoherent light and a sample holder that can be tilted permits three-dimensional ultrastructural imaging of cryopreserved adherent mammalian cells without chemical fixation.

    • Gerd Schneider
    • , Peter Guttmann
    •  & James G McNally

  • Research Highlights |

    The self-reconstructing properties of Bessel beams provide healing benefits in highly scattering media.

    • Daniel Evanko

  • Research Highlights |

    A noise-reduction algorithm decreases the amount of light needed for live-cell fluorescence microscopy by orders of magnitude.

    • Monya Baker

  • Research Highlights |

    Harmonic-generation microscopy takes the stage to provide biologists with a useful adjunct to fluorescence imaging.

    • Daniel Evanko

  • Brief Communication |

    Adaptations to total internal reflection microscopy permit visualization of the 'footprint' of rolling cells. Applied to neutrophils rolling in whole blood at physiological levels of shear stress, this approach reveals previously unappreciated features of rolling cell biology.

    • Prithu Sundd
    • , Edgar Gutierrez
    •  & Klaus Ley

  • Article |

    Generalized phase contrast and temporal focusing are combined to shape two-photon excitation patterns that elicit large photocurrents in ChR2-expressing neurons in culture and slices. This method allows precise aiming of the stimulating light at single neuronal processes, neurons or groups of neurons and can elicit simultaneous excitation of multiple cells using optogenetics.

    • Eirini Papagiakoumou
    • , Francesca Anselmi
    •  & Valentina Emiliani

  • Research Highlights |

    Visualization of newly synthesized proteomes using conventional fluorescence microscopy reveals the local nature of protein translation.

    • Erika Pastrana

  • Brief Communication |

    A monomeric fluorescent protein that can be irreversibly photoswitched from green to red form, both of which can be reversibly photoactivated, is reported. It is applied to pulse-chase experiments in which dynamic structures in live cells are imaged with superresolution using photoactivation localization microscopy (PALM).

    • Jochen Fuchs
    • , Susan Böhme
    •  & G Ulrich Nienhaus

  • Article |

    The combination of digital scanned laser light sheet microscopy and incoherent structured illumination allows intrinsic removal of scattered background fluorescence from the desired fluorescent signal. This provides substantial advantages for imaging nontransparent organisms and allowed reconstruction of a fly digital embryo from a developing Drosophila embryo.

    • Philipp J Keller
    • , Annette D Schmidt
    •  & Ernst H K Stelzer

  • News & Views |

    With a combination of microscopic and computational methods, the lineage of cells produced by divisions in the meristems of growing plants can now be tracked over time.

    • Edgar P Spalding

  • News & Views |

    New technical feats make it possible to monitor the activity of identified neurons in awake behaving Drosophila melanogaster.

    • Damon A Clark
    • , Saskia E J de Vries
    •  & Thomas R Clandinin

  • Article |

    This technique allows functional imaging of neurons in head-fixed Drosophila while the fly walks on an air-supported ball. Using a genetically encoded calcium sensor, the activity of motion-sensitive neurons in the fly optic lobe was recorded while the flies were presented with visual stimuli. Activity in these cells correlated with robust optomotor behavior in the walking flies.

    • Johannes D Seelig
    • , M Eugenia Chiappe
    •  & Vivek Jayaraman

  • Research Highlights |

    Multiple photodiode detectors are used to track the transit of dye-labeled single lipids through an excitation spot at high resolution.

    • Natalie de Souza

  • News & Views |

    Improved and easy-to-implement methods for precise fitting of curves to Poisson-distributed data—such as photons from single emitters—reach the limits of fitting precision.

    • Daniel R Larson

  • Article |

    Limitations in scanning speed have made it difficult for two-photon imaging to provide accurate temporal information on neuronal signaling. Refinements to random-access scanning using acousto-optic deflectors and an automated algorithm for reconstructing complex spike trains allowed in vivo high-speed optical recording of spiking activity in neuronal populations in the mouse neocortex.

    • Benjamin F Grewe
    • , Dominik Langer
    •  & Fritjof Helmchen

  • Brief Communication |

    A theoretical and experimental treatment of fitting methods for localizing the centers of diffraction-limited spots is presented. Use of an analytical point spread function shows that maximum likelihood fitting is superior to both unweighted and weighted least squares Gaussian fitting.

    • Kim I Mortensen
    • , L Stirling Churchman
    •  & Henrik Flyvbjerg

  • Technology Feature |

    Nonlinear optical microscopy lets researchers see chemical composition in living cells and organisms.

    • Monya Baker

  • This Month |

    Researchers watch proteins interact in real time.

    • Monya Baker

  • News & Views |

    A surprisingly simple method provides an effective way of correcting optical distortions in two-photon fluorescence microscopy and recovers nearly ideal images of inhomogeneous thick samples.

    • Rainer Heintzmann

  • This Month |

    Adapting optics: techniques for seeing stars scale to cells.

    • Monya Baker

  • Research Highlights |

    A miniature head-mounted two-photon microscope small enough for a rat to carry allows researchers to visualize neuronal signaling while the animal freely interacts with its environment.

    • Daniel Evanko

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