LSM 880 with Airyscan

ZEISS LSM 880 with Airyscan

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LSM 880 with Airyscan

Your New Standard for Fast and Gentle Confocal Imaging

Drosophila embryo. Color coded maximum intensity projection of the central nervous system.

Drosophila embryo. Color coded maximum intensity projection of the central nervous system. Courtesy of J. Sellin, AG Hoch, LIMES Institute, Bonn, Germany.

Drosophila embryo. Color coded maximum intensity projection of the central nervous system.

Drosophila embryo. Color coded maximum intensity projection of the central nervous system. Courtesy of J. Sellin, AG Hoch, LIMES Institute, Bonn, Germany

To get ahead in your research you may want to image the smallest structures, catch the faintest signals or track the fastest processes – or do all of that at once. When it comes to getting accurate data from live cells or other weakly-labeled samples, there is no such thing as too much sensitivity, resolution or speed. Each photon of emission light is precious.

With Airyscan you have the unrivaled combination of fast superresolution and sensitive confocal image acquisition at hand. Use multicolor samples with any label and get image quality like you’ve never seen before. Decide on this novel detector design and get a 4-8× improvement in signal-to-noise ratio (SNR) while simultaneously achieving a 1.7x increase in resolution. You will also be harnessing this unique combination of greater SNR and resolution at increased acquisition speeds of 27fps (at 480 x 480 pixels).

Highlights

Enter a New World of Confocal Performance with Airyscan

  • Gain sensitivity, resolution and speed for all your experiments. Image with negligible phototoxicity or bleaching - without changing your workflow, sample labels or system operation.
  • Airyscan’s unique Fast mode speeds up your imaging by a factor of four. This will propel you into the traditional domain of resonant scanning confocals without sacrificing sensitivity or resolution.
  • Airyscan delivers superresolution with high sensitivity at 140 nm laterally and 400 nm axially. This transcends the deconvolution approach by preserving precious emission light normally rejected at a closed pinhole.

 

 

Resonance Scanner using standard confocal

 

 

LSM 880 Airyscan Fast

Videos: Cardiomyocyte Cells with tubulin-EMTB to measure microtubule buckling. Left video: Resonance Scanner using standard confocal; Acquisition speed: 80fps. Right video: LSM 880 Airyscan Fast; Acquisitionspeed: 96 fps. Images and Samples courtesy of Ben Prosser, University of Pennsylvania, USA “Detyrosinated microtubules buckle and bear load in contracting cardiomyocytes”, Science April 2016.


HeLa cells
HeLa cells

Enhanced resolution with Airyscan. HeLa cells, Courtesy of S. Traikov, BIOTEC, TU Dresden, Germany

Enhanced resolution with Airyscan. HeLa cells, Courtesy of S. Traikov, BIOTEC, TU Dresden, Germany

 

Sample: U2OS Cells with Lifeactin; Max projection of 6 plane z-stack, 50 volumes over 17.5 minutes.


The patented Airyscan detector design allows gentle live cell imaging with superresolution. You collect more photons than with the closed pinhole of a traditional LSM and deconvolution. 

Increase Your Productivity

  • Combine the Fast mode for Airyscan with Z-stacks and tiling to image even large sample volumes with high image quality.
  • Collect all your fluorescent signals in one go. Parallel acquisition lets you investigate multiple labels in minimum time, equipped with the highest number of detectors of any confocal.
  • Improve your image quality with the unique combination of parallel spectral acquisition and high speed GPU deconvolution.
  • Make the most of large fields of view and the highest speed of any linear scanning confocal – ZEISS LSM 880 with Airyscan delivers up to 27 fps at 480x480 pixels in Fast mode.

 

 

Drosophila embryo, depth coded maximum intensity projection. Airyscan in Fast mode. Courtesy of B. Erdi, Max F. Perutz Laboratories, University of Vienna, Austria

 

Confocal Drosophila Brain NDD Airyscan.
Confocal Drosophila Brain NDD Airyscan.

Combine the resolution and sensitivity advantage of Airyscan with multiphoton excitation for superior imaging deep in your specimens. Confocal Drosophila Brain Zoom NDD Airyscan.

Confocal Drosophila Brain NDD Airyscan. 

 

  • You can use high frame rates in large fields of view with lowest light exposure for live imaging of whole organisms. 
  • Combine the resolution and sensitivity advantage of Airyscan with multiphoton excitation for superior imaging deep in your specimens. 

 

  • Combine the resolution and sensitivity advantage of Airyscan with multiphoton excitation for superior imaging deep in your specimens. 
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Choose the Most Flexible Confocal

  • Tailor Airyscan to your research needs with a choice of Superresolution, Sensitivity mode or the new Fast mode.
  • Remove autofluorescence and simultaneously separate highly overlapping fluorophores in a single scan. This will minimize phototoxicity in the sample.
  • Combine confocal imaging with single molecule techniques to gain mobility/concentration/oligomeric state information (FCS/FCCS/RICS/PCH).
  • Select Airyscan’s Fast mode to perform functional multiphoton imaging deep inside your sample.

 

Choanoflagellate rosette colony.

Choanoflagellate rosette colony. Blue: nuclei stained with Hoechst; magenta: tubulin of the flagella and cell body; green: actin microvilli collars stained with phalloidin. Courtesy of H. Aaron, UC Berkeley, USA

Choanoflagellate rosette colony.

Choanoflagellate rosette colony. Blue: nuclei stained with Hoechst; magenta: tubulin of the flagella and cell body; green: actin microvilli collars stained with phalloidin. Courtesy of H. Aaron, UC Berkeley, USA

 

Mitosis in HeLa-Kyoto cell line, imaged with ZEISS LSM 880 with Airyscan. Video showing Histone 2B (H2B, red, mCherry) and microtubule end binding protein 3 (EB3, green, EGFP) during mitosis.

  • Combine your data from multiple complementary imaging modalities – e.g., widefield, confocal and three different superresolution techniques. You can even correlate data with electron microcopy.
  • Gentle superresolution imaging allows long-term live cell imaging without artefacts caused by influencing the sample’s viability and biological function. 

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.

 

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.

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

See it for yourself!

Standard Confocal - 15 FPS; Pixel Dwell 320ns; Effective Laser Dosage 2.2x Airyscan Fast Mode - 30 FPS; Pixel Dwell 580ns; Color Coded Time Projection

Improve your highly dynamic Ca2+ imaging with the Fast mode for Airyscan. You capture more signal at high speeds while keeping light exposure to a minimum. Airyscan in Fast mode for Calcium Spark Imaging. With Airyscan in Fast superresolution mode you get more structural information at high speeds.
The donut shape of the Golgi in plant cells is nicely resolved. 

 

 

Use the white bar to slide between a standard confocal and the Airyscan Fast Mode. 

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. You can 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.

 

Applications & Videos

 

HeLa cells stained for Actin (green), Adapter Protein AP-3 (magenta) and Septin A (red). Courtesy of S. Traikov, BIOTEC, TU Dresden, Germany

Scroll through the Playlist to discover Airyscan Technology.

IMR90 human diploid lung fibroblasts.

IMR90 human diploid lung fibroblasts. Sample: Courtesy of J. Karlseder, Molecular and Cell Biology Laboratory; J. Fitzpatrick, Waitt Advanced Biophotonics Core, Salk Institute for Biological Studies, La Jolla, USA.

IMR90 human diploid lung fibroblasts.

IMR90 human diploid lung fibroblasts. DNA has been stained with DAPI, the telomeric G strand (leading strand) in green with a Peptide Nucleic Acid probe and Alexa 488 and the telomeric C strand (lagging strand) in red with a Peptide Nucleic Acid probe and Alexa 546. Prior to their harvest the cells have been treated with siRNAs targeting RTEL1. RTEL1 is a helicase that is essential for telomere replication, and lack of the protein leads to stalled forks at telomeres and telomere breakage. This can be seen by individual telomeres that appear as more than one dot, as highlighted in the images. Airyscan resolves multiple telomere dots, thereby allowing an accurate quantification of telomere replication problems. Sample: Courtesy of J. Karlseder, Molecular and Cell Biology Laboratory; J. Fitzpatrick, Waitt Advanced Biophotonics Core, Salk Institute for Biological Studies, La Jolla, USA.

Human RPE cells

Human RPE cells, ZO1 (tight junction marker) in blue, photoreceptor outer segments stained with FITC in green, EEA1 (endosomal marker) in red. Courtesy of S. Almewadar, CRTD, TU Dresden, Germany

Human RPE cells

Human RPE cells, ZO1 (tight junction marker) in blue, photoreceptor outer segments stained with FITC in green, EEA1 (endosomal marker) in red. Courtesy of S. Almewadar, CRTD, TU Dresden, Germany

 

Fixed tumor cells, tubulin labelled with Alexa 555, Airyscan SR mode. Sample courtesy of P. O'Toole and P. Pryor, University of York, UK. 

 

Oligodendrocyte, CNPase-antibody staining. Courtesy of C. Dornblut, Leibniz Institute for Age Research (FLI), Jena, Germany

 

Drosophila embryo, maximum intensity projection. Microtubules labelled with GFP. Left: z-stack with 55 slices. Imaged for 203 min at 3 min interval. Courtesy of B. Erdi, Max F. Perutz Laboratories, University of Vienna, Austria

 

Drosophila embryo, maximum intensity projection. Microtubules labelled with GFP. Imaged at higher magnification. Z-stack with 117 slices, imaged for 75 min at 3 min interval. Courtesy of B. Erdi, Max F. Perutz Laboratories, University of Vienna, Austria

Watch the Product Trailer

 

Downloads

ZEISS LSM 880 with Airyscan

Your New Standard for Fast and Gentle Confocal Imaging

Pages: 36
Filesize: 11,605 kB

White Paper: Airyscanning - Short version

A Novel Approach to Confocal Imaging

Pages: 5
Filesize: 1,709 kB

Beampath - ZEISS LSM 880 with Airyscan

Revolutionize Your Confocal Imaging

Pages: 1
Filesize: 607 kB

Application Note: Cryo-Confocal Imaging with Airyscan

Improving Resolution and Signal-to-Noise in Cryo-Fluorescence Microscopy

Pages: 8
Filesize: 1,483 kB

Technology Note: The Airyscan Detector from ZEISS

Confocal Imaging with Improved Signal-to-Noise Ratio and Superresolution

Pages: 19
Filesize: 3,958 kB

Application Note: Spectral Imaging: a Powerful Tool for Confocal Multicolor Imaging in Living Plant Cells

Pages: 7
Filesize: 1,138 kB

Technology Note: ZEISS LSM 880 with Airyscan

Introducing the Fast Acquisition Mode

Pages: 7
Filesize: 1,071 kB

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