Zeiss LSM 980-Airyscan2

Super-Resolution Using the Airyscan

  1. Can I really get better resolution using an Airyscan detector? This is a good introduction to the resolution obtainable using Huygens on Airyscan data.
    Huygen’s Array-detector (Airyscan®) Webinar from April 30th, 2019
  2. How does the Airyscan give you much better sensitivity? These figures will help you understand why an Airyscan detector gives you much better signal to noise in your image, which is key to obtaining super-resolution later.
  3.  But how does the Airyscan produce more resolution?Probably your best bet to understand the increased resolution with the Airyscan can be found here:  AIRY scan basic principle, particularly pages 7 and onward.  
    1. With the Airyscan you are creating a point-scanner detector with many of the characteristics of a wide-field experiment (higher spatial resolution and much, much better signal-to-noise).
    2. The Airyscan retains all the speed and sample penetration benefits of a point scanner
    3. While there is no classical pin hole in the 980-Airyscan2, the size of each of the 32 individual PMTs corresponds to a 0.2 AU pinhole. Although with a traditional confocal you would quickly fry your sample because of the lack of signal with a pinhole this small, the signal to noise boost with 32 concentric detectors pulls the extra resolution out of an otherwise noisy signal.
    4. Like with a wide-field experiment, the largest increases in resolution occur when you deconvolve a 3-D stack of Airyscan images
    5. It is possible to obtain a 1.7-fold improvement in all spatial directions.  For emitted light at 480 nm, this corresponds x,y resolution of 140 nm and z resolution of 400 nm.
    6. Airyscan images contain high spatial frequency (fine detail) information as high/fine as SIM, but the proportion of the image intensity with high resolution is smaller with the Airyscan than with the SIM.  This is similar to Michael’s experience with the Nikon N-SIM relative to our 980-Airyscan2.