Comparing Airyscan technology to conventional confocal imaging in live cell imaging

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The recently published peer-reviewed paper “Exploring the Potential of Airyscan Microscopy for Live Cell Imaging” by Kseniya Korobchevskaya, B. Christoffer Lagerholm, Huw Colin-York and Marco Fritzsche from the University of Oxford, UK highlights the significant improvement of resolution and signal-to-noise at the same time of ZEISS Airyscan compared to conventional confocal imaging techniques. The article was published as part of the Special Issue Superresolution Optical Microscopy.

“Biomedical research demands  non-invasive and ultra-sensitive imaging techniques. Especially, our laboratory for Biophysical immunology at the MRC Human Immunology Unit and Kennedy Institute for Rheumatology at the University of Oxford relies on state-of-the-art imaging technology with extended spatial and temporal resolution as offered by the novel ZEISS Airyscan technology. In our recent  paper, we demonstrate how Airyscan imaging successfully bridges the gap between conventional confocal and super-resolution microscopy”, says  Marco Fritzsche.

Abstract: Biological research increasingly demands the use of non-invasive and ultra-sensitive imaging techniques. The Airyscan technology was recently developed to bridge the gap between conventional confocal and super-resolution microscopy. This technique combines confocal imaging with a 0.2 Airy Unit pinhole, deconvolution and the pixel-reassignment principle in order to enhance both the spatial resolution and signal-to-noise-ratio without increasing the excitation power and acquisition time. Here, we present a detailed study evaluating the performance of Airyscan as compared to confocal microscopy by imaging a variety of reference samples and biological specimens with different acquisition and processing parameters. We found that the processed Airyscan images at default deconvolution settings have a spatial resolution similar to that of conventional confocal imaging with a pinhole setting of 0.2 Airy Units, but with a significantly improved signal-to-noise-ratio. Further gains in the spatial resolution could be achieved by the use of enhanced deconvolution filter settings, but at a steady loss in the signal-to-noise ratio, which at more extreme settings resulted in significant data loss and image distortion.

Time-lapse images of activating Rat Basophilic Leukaemia (RBL) cell. (a) Comparison of 1.25 AU confocal and Airyscan processed AF6.7 images. Scale bar is 10 µm. (b) Direct comparison of Region of Interest area (ROI; red rectangle in (a)) between confocal 1.25 AU and Airy processed images with AF4, AF6 and AF7, respectively. Scale bar is 5 µm. (c) Time-lapse of activating RBL cell at 0, 60 and 120 s, respectively. Green LifeAct-citrine (excitation at 488 nm), red SNAP-tag (excitation at 561 nm). (d) Intensity profiles from 1.25 AU (grey filled), AF7 (blue dots) and AF8 (red) images along the line indicated by white arrows in (b). Arrows indicate peaks from two separate actin fibres, which are only distinguishable at high AF strength and are not resolved at 1.25 AU. Korobchevskaya K, Lagerholm BC, Colin-York H, and Fritzsche M, Exploring the Potential of Airyscan Microscopy for Live Cell Imaging, Photonics, 2017.

The authors of this paper focused on the superresolution aspect. To learn more about the enhanced sensitivity and speed of ZEISS Airyscan read this free paper.

Watch this video and understand how ZEISS Airyscan works:

 

Read the full paper: Korobchevskaya K, Lagerholm BC, Colin-York H, and Fritzsche M, Exploring the Potential of Airyscan Microscopy for Live Cell Imaging, Photonics, 2017

More information on ZEISS Airyscan

Tags: Airyscan, Confocal Microscopy

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