The technology behind it

LSM 880 with Airyscan

The Airyscan Principle

The Airyscan Principle

The Airyscan Principle - Revolutionizing your confocal imaging: the beam path of LSM 880 with Airyscan and the Fast mode.

The Airyscan Principle

The Airyscan Principle - Revolutionizing your confocal imaging: the beam path of LSM 880 with Airyscan and the Fast mode

LSM 880 with Airyscan Beampath - Revolutonize Your Confocal Imaging

 

Airyscan is a detector that draws on the fact that a fluorescence microscope will image a point-like source as an extended Airy disk (Airy pattern). In a standard confocal microscope the out-of-focus emission light is rejected at a pinhole, the size of which determines how much of the Airy pattern reaches the detector. In a standard confocal microscope, you get a sharper image by gradually closing the pinhole to reject out-of-focus light. However, it’s also dimmer since a great deal of light is then lost. The smaller the pinhole, the higher the resolution, but – equally – the greater the loss in light.

Airyscan solves this conundrum between resolution and light efficiency by imaging the Airy disk onto a concentrically-arranged hexagonal detector array. Its detection area consists of 32 single detector elements, all of which act like very small pinholes. The confocal pinhole itself remains open and doesn’t block light – thus all photons of the whole Airy disk are collected.

The signals from all detector elements are then reassigned to their correct position, producing an image with increased signal-to-noise ratio and resolution.

An area detector consisting of multiple detector elements allows great flexibility in imaging modes. In Fast mode, the excitation beam is elongated in y and, with one horizontal scanner movement, the Airyscan detector acquires four lines of image information instead of only one. This parallelization delivers a unique combination of high speed, high resolution and high sensitivity. Airyscan capitalizes on the scanning and optical sectioning capabilities of a confocal and therefore works with your standard samples, standard dyes and even with thicker samples such as tissue sections or whole animal mounts that need a higher penetration depth.

It's up to you whether you use the strengths of Airyscan and its unique Fast mode to get better signal-to-noise, superresolution or speed. With single or multiphoton excitation.

With Airyscan in Fast superresolution mode you get more structural information at high speeds.

Comparison of SR-SimM (left) and Airyscan (right). The robust optical sectioning capability of Airyscan allows superresolution imaging even in thicker samples. Stereo cilia in vestibular hair cells. Sample courtesy of Drs. Matt Avenarius, Jocelyn Krey and Peter Barr-Gillespie, OHSU.

Airyscan. Stereo cilia in vestibular hair cells. (Same sample as on the left hand side)

The donut shape of the Golgi in plant cells is nicely resolved. Arabidopsis thaliana root. First image: LSM, GaAsP; second image: Fast mode, SR; third image: Airyscan, SR. Samples provided by Professor C. Hawes, Oxford Brookes University, UK.

The donut shape of the Golgi in plant cells is nicely resolved. Arabidopsis thaliana root. First image: LSM, GaAsP; second image: Fast mode, SR; third image: Airyscan, SR. Samples provided by Professor C. Hawes, Oxford Brookes University, UK.

The donut shape of the Golgi in plant cells is nicely resolved. Arabidopsis thaliana root. First image: LSM, GaAsP; second image: Fast mode, SR; third image: Airyscan, SR. Samples provided by Professor C. Hawes, Oxford Brookes University, UK.

Arabidopsis thaliana root. First image: LSM, GaAsP; second image: Fast mode, SR; third image: Airyscan, SR. Samples provided by Professor C. Hawes, Oxford Brookes University, UK.

Your Powerful Combination

Count on LSM 880, the fastest linear scanning confocal on the market, to fully resolve the movement of labeled proteins in dynamic cellular and subcellular processes.

Add Airyscan with the Fast module to image with up to 27 frames per second at 480 × 480 pixels with highest image quality. LSM 880 is constantly monitoring and calibrating the scanner position to guarantee a stable field of view and equal pixel integration times over the whole field of view. Linear scanning is aprerequisite for both gentle image acquisition, and quantitative and correlative imaging. It gives you a constant signal-to-noise level and uniform exposure to the illuminating laser throughout the scanned area, including manipulated regions of interest. Unlike traditional sine scanning confocals, LSM 880 uses more than 80% of the scanning time for data acquisition. Longer pixel integration times at a defined frame with a 29% better signal-to-noise ratio give you the best image quality, even for your most demanding samples.

 

Capture more information by using the Fast mode for Airyscan for rapid volume imaging of dynamic processes. With multiphoton excitation you observe Ca 2+ dynamics in 3D. Calcium imaging of Zebra fish spine. GCaMP5, 920 nm excitation, 9 z-slices over 18 μm. Airyscan in Fast NLO mode. Sample: Courtesy of D. Friedmann, UC Berkeley, USA.

 

 

With Airyscan in Fast mode you obtain more structural information at high speeds. Endoplasmic reticulum labelled with p35S::GFP-HDEL, Golgi p35S::ST-RFP, stably transformed A. thaliana cells. Root Time SR Fast Airyscan Processing. Courtesy of Professor C. Hawes, Oxford Brookes University, UK.

 

High sensitivity and low phototoxicity for live imaging of delicate samples. C. elegans Gap Junction imaging with the Fast mode for Airyscan.C. elegans embryo. Adherens junction protein labelled with GFP. Maximum intensity projection of a z-stack with 100 slices. Imaged for 120 min at 5 min interval. Courtesy of L. Cochella, Research Institute of Molecular Pathology (IMP), Vienna, Austria.

 

Parallel Acquisition of Multiple Channels

Airyscan Beampath from ZEISS LSM 880 with Airyscan

Airyscan Beampath from ZEISS LSM 880 with Airyscan.

Airyscan Beampath from ZEISS LSM 880 with Airyscan

Airyscan Beampath from ZEISS LSM 880 with Airyscan

It takes multiple labels to analyze interactions between different cellular or subcellular structures. Gain new insights into biological structures and processes by acquiring the entire fluorescent spectra of all your labels at once. LSM 880 delivers 32 channels with 10 nm spectral resolution and 512 x 512 pixels at 5 fps.

Set up 10 channels for multichannel spectral imaging and then add the transmission detector. You can now image all fluorescent dyes and the additional oblique contrast in a single scan. This protects your sample and saves time, too.

Especially for those demanding multiphoton experiments, you have the advantage of this fundamental capability: up to 12 non descanned detectors can be read out in parallel.

 

Acquire the whole fluorescence spectra of your sample gently with one single scan and separate all your fluorescent labels and autofluorescence. 

Time-lapse movie of the actin filaments (green), endoplasmic reticulum (magenta), MitoTracker Orange (yellow), and chlorophyll auto-fluorescence (blue). Images were taken every two seconds for 4 min. 30× real time.Courtesy of Takehiko Kanazawa and Takashi Ueda, Department of Biological Sciences, Graduate School of Science, University of Tokyo; Atsuko Era, Center for Frontier Research, National Institute of Genetics.

 

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