ZEISS LSM 980 with Airyscan 2

Your Next Generation Confocal Microscope with Multiplex Mode

Your life sciences research can be demanding, and if you are involved in neuroscience, cancer research or other cell- or organism-based disciplines, you’ll often need confocal microscopy data for your work. Emerging technologies such as CRISPR / Cas open up innovative ways of thinking and allow you to ask altogether new scientific questions, deeply affecting your imaging experiments. To monitor life as undisturbed as possible requires low labeling density for your biological models – for example, 3D cell culture, spheroids, organoids or even whole organisms – and this calls for 3D live cell imaging that combines optical sectioning with low phototoxicity and high speed. Then there are the repeated experiment runs it takes to get statistically valid data for your conclusions: it soon becomes apparent you will also need high throughput.

Your new LSM 980 with Airyscan 2 is the ideal confocal microscope for 4D imaging. The entire beam path is optimized for simultaneous spectral detection of multiple weak labels with the highest light efficiency. Add the new Multiplex mode for Airyscan 2 to get more imaging options for your experiments. You can now choose the perfect setup to gently image larger fields of view with superresolution in shorter acquisition times than ever before. A number of software helpers will optimize your workflow and support efficient acquisition and data management. With ZEN Connect you can document and share all details of your experiments. You’ll always keep the context as you combine overview images, ROIs and additional data, even across imaging modalities.

Highlights

  • Get Better Data Faster

Use the new Multiplex mode for Airyscan 2 and get more information in less time. Smart illumination and detection schemes let you image your most challenging three-dimensional samples with high framerates beyond the diffraction limit and still treat your sensitive samples gently. By combining the full flexibility of a point scanning confocal with the speed and gentleness of the sensitive Airyscan area detector, it’s now possible to answer your scientific questions eight times faster with superresolution.




 

Live imaging with 143 frames per second of fluorescently labeled motile cilia of brain ependyma. Acquired with Airyscan CO-8Y mode combining image quality and speed; for detailed analysis of ciliary beating direction and frequency. © Courtesy of G. Eichele, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

  • Increase Your Productivity
It’s never been easier to set up complex live cell imaging experiments on a confocal microscope. The latest version of ZEN imaging software drives your new LSM 980 with Airyscan 2 and puts a wealth of software helpers at your command. You’ll work easier than ever before and faster, too, achieving reproducible results in the shortest possible time. Smart Setup and the new Sample Navigator let you find and image regions of interest quickly, leaving more time for the real work of acquiring data. Direct Processing enables parallel acquisition and data processing. ZEN Connect keeps you on top of everything, both during imaging and later when sharing the whole story of your experiment. It’s easy to overlay and organize images from any source.

See how ZEN Connect helps to always keep your context while imaging. From acquiring an overview image, to defining ROI's, and
even when changing between different imaging systems. You save time and always stay on top of things.

  • Image with More Sensitivity

LSM 980 brings you the best of two worlds to image your most challenging samples. You get a confocal microscope with the light efficient beam path of the LSM 9 family with up to 34 simultaneous channels for full spectral flexibility. This lets you image faint signals with the highest sensitivity. Plus, you can combine this confocal microscope with the new Multiplex mode for Airyscan 2. This revolutionary method advances confocal microscopy and extracts even more information from your sample in less time. You don't need to close a pinhole to get superresolution, which makes your confocal 4D imaging even more light efficient. Expect the very best data quality from all your samples.

 

HeLa cells stained for DNA (blue, Hoechst 44432), microtubules (yellow, anti-tubulin Alexa 488) and F-actin (magenta, phalloidin Abberior STAR Red).

HeLa cells stained for DNA (blue, Hoechst 44432), microtubules (yellow, anti-tubulin Alexa 488) and F-actin (magenta, phalloidin Abberior STAR Red). Imaged with ZEISS Airyscan 2 in Multiplex mode for efficient superresolution imaging of a large field of view. © Courtesy of A. Politi, J. Jakobi and P. Lenart, MPI for Biophysical Chemistry, Göttingen, Germany.

A Flexible and Sensitive Confocal Microscope

For Best Image Quality

The beam path design and every single component of your confocal microscope LSM 980 with Airyscan 2 are optimized to deliver the highest sensitivity and flexibility and up to 13 frames / sec for your experiments. Linear scanners illuminate your sample evenly for efficient signal collection during more than 80 % of the frame time. The special low angle orientation of the Twin Gate main dichroic beam splitter suppresses stray light and always gives you crisp contrast. You can even extend your emission detection range over the excitation laser line to make sure you collect all of those precious emission photons.

Your LSM 980 directs emission light to the 3, 6 or 34 channel Quasar detector where all signals from your dye combination are measured. Match the emission detection bands to your labels with nanometer precision. You can acquire overlapping labels or autofluorescence in a single lambda scan with 34 channels and then separate them with Linear Unmixing. You keep your sample’s light exposure to a minimum and speed up your confocal imaging. With LSM 980, you use the enhanced quantum efficiency and excellent dynamic range of sensitive GaAsP detectors. Then add Airyscan 2 for enhanced sensitivity, speed and superresolution – and combine all imaging modes into one single experiment.

The Airyscan Principle

Principle of airyscan

Classic confocal laser scanning microscopes use point illumination to scan the sample sequentially. The microscope optics transform each point to an extended Airy disk (Airy pattern). A pinhole then spatially limits this Airy disk to block out-of-focus light from reaching the detector. Closing the pinhole gives higher resolution, but at the price of detecting fewer photons – and these photons cannot be brought back by e.g. deconvolution.

Airyscan 2 is an area detector with 32 concentrically arranged detection elements. This allows you to acquire more of the Airy disk at once. The confocal pinhole itself remains open and does not block light, thus more photons are collected. This produces much greater light efficiency while imaging. Airyscan 2 gives you a unique combination of gentle superresolution imaging and high sensitivity.

How the New Multiplex Mode for Airyscan 2 Works

The LSM 9 family with Airyscan 2 from ZEISS now gives you more options to fit imaging speeds and resolution to your experimental needs. You combine a confocal area detector with smart illumination and readout schemes, which let you choose from different parallelization options. The new Multiplex mode uses knowledge about the shape of the excitation laser spot and the location of single area detector elements to extract more spatial information, even during parallel pixel readout. This allows taking bigger steps when sweeping the excitation laser over the field of view, improving your achievable acquisition speeds.

In fact, the high amount of spatial information captured in the pinhole plane allows reconstructing a final image with better resolution than the acquisition sampling. Airyscan 2 in Multiplex mode can acquire up to four superresolution image lines with high SNR in a single sweep. Your LSM 980 with Airyscan 2 allows to stretch the excitation laser spot to image eight lines in parallel. Use this speed advantage for ultrafast time series of single slices, for rapid tiling of large areas or for fast volumetric time-lapse imaging.

Watch the Multiplex mode animation trailer

LSM 980 Airyscan SR Multiplex SR-4Y Multiplex SR-8Y Multiplex CO-8Y
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Max. fps at max. field of view

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9.6 (Zoom 1)

Antibody labelling, fine structures

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Antibody labelling, tiling

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Live cell imaging

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Applications

ZEISS LSM 980 at Work

Meiosis in starfish oocytes
The depth coding shows a subset of 52 μm. The movie shows the transport of chromosomes, labeled by Histone 1-Alexa 568, in a starfish oocyte undergoing meiosis. A z-stack of 67 μm was acquired every 2.4 seconds with Airyscan CO-8Y mode. Concomitant with chromosome transport, the nucleolus (the large spherical structure) is disassembling.

Meiosis in starfish oocytes
The rendering is a projection of the process along z-axis (maximum intensity) and time (color-coded projection); to illustrate the movement of the chromosomes within the volume of the nucleus. © Courtesy of P. Lenart, MPI for Biophysical Chemistry, Göttingen, Germany.

Oocytes store all the nutrients to support early embryonic development, and are therefore very large cells with a large nucleus. Oocytes need to divide before fertilization. How to make cell division work in this very large cell is the topic investigated by P. Lenart’s lab.
They have shown that, surprisingly, an actin network is required to collect chromosomes scattered in the oocyte nucleus. They are then handed over to microtubules, which capture chromosomes and align them on the spindle. The actin-driven and microtubule-driven transport phases have very different speeds and show other differentiating characteristics that can be distinguished by tracking chromosome motion.

This is a nice imaging challenge, because chromosomes are scattered in the spherical nucleus with a diameter of 80 μm and are transported over a period of approximately 15 minutes. Back in 2005 we could acquire stacks every 45 s, which was sufficient to distinguish actin- and microtubule-driven phases. Using the new, high resolution trajectories shown here we hope to learn about the details of the transport mechanism.

Peter Lenart

tissue explant of ependyma from the ventricular system of a mouse brain

This ZEN Connect project documents the experiment performed with the tissue explant of ependyma from the ventricular system of a mouse brain. All acquired data of the experiment session is kept in context. The overview images by camera and LSM allow to precisely record the localization of the acquired ciliary beating within the sample. The flow map of cilia generated flow along the ependymal wall is added as a reference.

overview of fluorescently labeled motile cilia on ependyma tissue explant from the mouse brain

An overview of fluorescently labeled motile cilia on ependyma tissue explant from the mouse brain is quickly acquired by tiling with Airyscan 2 in Multiplex CO-8Y mode to find regions of interest. Z-Stack displayed in colored depth coding. The exact position of the recorded motile cilia is documented.

Live imaging with 143 frames per second of fluorescently labeled motile cilia of brain ependyma. Acquired with Airyscan CO-8Y mode combining image quality and speed; for detailed analysis of ciliary beating direction and frequency. © Courtesy of G. Eichele, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.


© Courtesy of M. Paoli, Galizia Lab, University of Konstanz, Germany.

The brain, thoracic and abdominal ganglia of the cockroach are joined together by bilateral connective bundles of ascending and descending interneurons forming the ventral nerve cord. In this preparation, left and right connectives were individually labelled (Alexa 488: green, Alexa 647: magenta) posteriorly to the suboesophageal ganglion to observe the extension of their innervation within the different neurophils, and throughout the ipsi- and contralateral parts of the brain (DNA labelled with DAPI: cyan). Imaging was performed using Tiling and Stitching to capture the complete volume (3×2.3× 0.26 mm). 3D animation of the complete dataset was done with arivis Vision 4D, ideal for rendering and analyzing large datasets. The 4D viewer in arivis Vision 4D can be configured to adjust the appearance of individual channels independently to highlight specific features.

Theses settings, along with clipping planes or the varying opacity of individual channels, can be stored into key frames which the software automatically interpolates between to produce a seamless animation. These animations can be previewed and edited prior to producing high resolution video renders.

Downloads

ZEISS LSM 980 with Airyscan 2

Your Next Generation Confocal for Fast and Gentle Multiplex Imaging

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Filesize: 10,887 kB

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