Your New Standard for Fast and Gentle Confocal Imaging
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To get ahead in your research you may want to image the smallest structures, catch the faintest signal 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 for this novel detector design and get a 4-8× improvement in signal-to-noise ratio (SNR) as compared to imaging with conventional confocal GaAsP detectors. And 1.7× higher resolution for your single photon or multiphoton experiments.
Heterochromatin protein 1 (HP-1) fused to GFP and expressed in the nucleus of a human Hep G2 cell. Left panel shows the distribution of HP-1 between Euchromatin and denser Heterochromatin areas. Right panel represents a brightness map demonstrating dimerization of HP-1 within heterochomatic regions. Sample courtesy of P. Hemmerich, Leibniz-Institute for Age Research (FLI), Jena, Germany.
Mitochondria, RK 13 TOMM20 labeled with
Cerulean 3; comparing confocal GaAsP and
Airyscan detection. Sample: Courtesy of
M. Davidson, The Florida State University,
With Airyscan You Enter a New World of Confocal Performance
Perform Quantitative Imaging
Increase Your Productivity
LSM 880 with Airyscan Beampath -
Revolutonize Your Confocal Imaging
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)
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. When you increasingly close the pinhole in a standard confocal microscope to reject out-of-focus light, you get a sharper image, but it’s also dimmer since a great deal of light is then lost. The smaller the pinhole, the higher the resolution, but – equally – the bigger 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 for great flexibility in imaging modes. In Fast mode, the excitation beam is elongated in y and the Airyscan detector acquires 4 lines of image information instead of only one with one horizontal scanner movement. 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 advantages of Airyscan and the Fast module to get better signal-to-noise, superresolution or speed – with single or multiphoton excitation.
African green monkey kidney. TOMM20
(Alexa 568, red), Tubulin (Alexa 488, green)
and nucleus (DAPI, blue). Sample: Courtesy
of M. Davidson, The Florida State University,
Arabidopsis root. Green: endoplasmatic
reticulum labelled with GFP, Magenta:
Golgi labelled with RFP. Courtesy of
C. Hawes, Oxford Brookes University, UK
To fully resolve the movement of labeled proteins in dynamic cellular and subcellular processes you often need to image at around 10 frames per second. Now, with LSM 880, you can achieve up to 13 frames per second at 512 × 512 pixels.
Add Airyscan with the Fast module to image with up to 27 frames per second at 480 × 480 pixels.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 an essential prerequisite for your 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. That means you will enjoy a 29% improvement of signal-to-noise ratio because of longer pixel integration times at a defined frame rate.
It takes multiple labels to analyze interactions between different cellular or subcellular structures, but you can achieve the highest timing precision and speed up your imaging time by recording their intensities simultaneously. LSM 880 lets you acquire the entire spectrum – and all your labels – in just one scan with 32 channels , 512 x 512 pixels at 5 frames per second.
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 also saves you time.
Especially for your demanding multiphoton experiments, you will profit from having this fundamental capability: up to 12 non descanned detectors can be read out in parallel.
HeLa cells stained for Actin (green), Adapter Protein AP-3 (magenta) and Septin A (red). Courtesy of S. Traikov, BIOTEC, TU Dresden, Germany
Drosophila embryo, depth coded maximum intensity projection. Airyscan in Fast mode. Courtesy of B. Erdi, Max F. Perutz Laboratories, University of Vienna, Austria
Calcium sparks labeled with Fluo 4 imaged in Cardiomyocytes with 50 frames per second. Airyscan in Fast mode. Courtesy of P. Robison, B. Prosser, University of Pennsylvania, USA
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.
Skin tissue from pig labelled with Ethyleneblue. The unfixed sample was excited with 1100 nm using an OPO (optical parametric oscillator). Fluorescent lifetime measurement was performed using the detector module BiG.2 connected to the TCSPC electronics from Becker&Hickl. The color coded image shows the variation in lifetime within different types of skin cells.
Oligodendrocyte, CNPase-antibody staining. Courtesy of C. Dornblut, Leibniz Institute for Age Research (FLI), Jena, Germany
Slice of mouse brain, CNPase-antibody staining, imaged with 10x objective. Courtesy of C. Dornblut, Leibniz Institute for Age Research (FLI), Jena, Germany
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.
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.
Arabidopsis root. Green: endoplasmatic reticulum labelled with GFP, magenta: Golgi labelled with RFP. Confocal GaAsP detection. Courtesy of C. Hawes, Oxford Brookes University, UK
Arabidopsis root. Green: endoplasmatic reticulum labelled with GFP, magenta: Golgi labelled with RFP. Airyscan in Fast mode. Courtesy of C. Hawes, Oxford Brookes University, UK
Arabidopsis root. Golgi labelled with RFP. Left: confocal GaAsP detection, middle: Airyscan in Fast mode, right: Airyscan in SR mode. Courtesy of C. Hawes, Oxford Brookes University, UK
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
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
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
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