ZEISS LSM 900 with Airyscan 2

Your Compact Confocal for Multiplex Imaging

What are you looking for in confocal imaging? Whatever your scientific question, you want to start with the best possible image quality and that means crisp contrast and the best resolution. You also want the highest sensitivity for gently imaging your living or fixed samples without bleaching. Your confocal LSM 900 with Airyscan 2 has all this and more. You image with 4 – 8× more signal-to-noise ratio (SNR) and with superresolution. You also get the highest frame rates: the new Multiplex mode for Airyscan 2 adds smart detection schemes for parallel pixel acquisition. You can now observe dynamic processes in living specimens gently – without sacrificing image quality. Plus, your LSM 900 has a genuinely small footprint, concentrating on the essence of a confocal and leaving off needless complexity. It fits easily into your lab or imaging facility – and it’s easy to use, too.

Highlights

  • Get Better Data Faster

Combine the excellent image quality of your confocal LSM 900 with the new Multiplex mode for Airyscan 2 to get more information in shorter times. You can now employ smart detection schemes to image your challenging three-dimensional samples with the highest framerates and with increased resolution of 140 nm. The speed and gentleness of the sensitive Airyscan area detector complement this compact point scanning confocal and allow you to image your most demanding samples with 4 – 8× more SNR.




HeLa cells stained for DNA (blue, Hoechst 44432), microtubules (green, anti-tubulin, alpaca anti-mouse

HeLa cells stained for DNA (blue, Hoechst 44432), microtubules (green, anti-tubulin, alpaca anti-mouse-alexa 488) and F-actin (red, phalloidin-Abberior STAR Red). Acquired with Multiplex mode for ZEISS Airyscan. © Courtesy of A. Politi, J. Jakobi and P. Lenart, MPI for Biophysical chemistry, Göttingen, Germany.

  • Increase Your Productivity

Your LSM 900 with Airyscan 2 is not just a compact confocal – it's also very easy to use. Setup is simple with ZEN imaging software, even for complex confocal live cell imaging experiments. A wealth of software helpers lighten your workload and make sure you get reproducible results in the shortest possible time. Smart Setup and the new Sample Navigator let you find regions of interest and image them quickly, leaving you more time to acquire data. Direct Processing allows parallel acquisition and data processing. ZEN Connect keeps you on top of things at all times, 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.

  • A Small Footprint for Greater Image Quality

Your LSM 900 is packed with innovative and clever solutions for producing the best quality in confocal live cell imaging. The elegant beam path is designed for high spectral flexibility, with each single component optimized for the highest sensitivity and contrast. Given its small footprint and reduced complexity, you'll save valuable lab space, minimize the time needed for user-training and reduce the cost of ownership – this is especially good news for imaging facilities.





Cell division of LLC-PK1 cells, alpha-tubulin (mEmerald, magenta) and H2B (mCherry, green). With the new Multiplex mode for ZEISS Airyscan a Z-stack of 52 slices was captured every 40 seconds for a total of 40 minutes.

A Streamlined Light Path

With Surprising Flexibility

The compact light path with a minimum of optical elements is designed for highest efficiency. Fluorescence emission light travels through the main dichroic beam splitter with its outstanding laser suppression to deliver supreme contrast. Up to two patented variable beam splitter dichroics (VSDs) divert the spectral portions of emission light. You can define up to three detectors - multialkali, GaAsP or Airyscan 2. GaAsP PMTs – that is, gallium arsenide phosphide photomultiplier tubes – display high light collection efficiencies over a broad spectral range. Their low dark noise levels also render them the ideal tool for detecting faint signals.

Enjoy outstanding image quality based on a superb signal-to-noise ratio (SNR). You might use this gain in SNR to increase productivity by achieving faster scan speeds while preserving excellent image quality. Or take advantage of the low laser powers needed in confocal live cell imaging applications to avoid photobleaching and phototoxicity. Or simply detect faint signals in low expressing cells. All that, and you can do it with up to three spectral confocal channels simultaneously.

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

Watch the Multiplex mode animation trailer

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.

LSM 900 Airyscan SR Multiplex SR-2Y Multiplex SR-4Y Multiplex CO-2Y
Parallelization

1

2

4

2

Resolution

120/120

140/140

140/140

Confocal or better

Max. fps at max. field of view

0.4

0.8

3.5

3.5

Antibody labelling, fine structures

+++++

++++

++++

++

Antibody labelling, tiling

++

+++

+++++

+++

Live cell imaging

++

+++

++++

+++++

Applications

ZEISS LSM 900 at Work

Drosophily embryo imaged with Multiplex Mode for LSM 900.
click to enlarge
COS 7 cells labelled microtubules Aairyscan-Modus

Drosophily embryo – imaged with Multiplex Mode for LSM 900.
© Courtesy of J. Sellin, LIMES, Bonn, Germany 

Comparing the field of view you can image at superresolution in the same time using Multiplex mode (left) and Airyscan SR (right). COS 7 cells with labelled microtubules (alpha-tubulin 488, green) and actin (phalloidin 568, red).  

Lateral line primordium migration and deposition of immature neuromasts in a Zebrafish embryo (Danio rerio).
Animals were anesthetized and embedded using low concentrated agarose in a glass bottom petridish.
Initial camera based imaging allowed for a quick and easy sample navigation (top) combining Phase Gradient Contrast with fluorescence acquisition. Subsequent high resolution imaging with Airyscan 2 in Multiplex mode was done on individual positions identified in the widefield image (white boxes).
The gentle and fast image acquisition that is inherent to the Airyscan 2 Multiplex mode is very beneficial for this kind of application. The animal is unperturbed by the imaging while images with a very high signal to noise ratio as well as level of detail can be acquired at the same time.

Green: LYN-eGFP (mebranes); Red: tagRFP-T-UTRCH (actin). © Courtesy of J. Hartmann and D. Gilmour, EMBL, Heidelberg, Germany


Human lung epithelial cell line A549 stained with MitotrackerOrange (mitochondria) and SIR-DNA (nuclei).

The acquisition seemlessly combines two imaging modes – the fluorescent channels where captured in confocal mode using highly sensitive GaAsP detectors while the Phase Gradient Contrast is camera based.

A timelapse of 2.5 h was acquired using a 40× magnification with a numerical aperture of 0.95.

© A.C. Hocke, Charité, Berlin, Germany.


Sometimes you need to see and assess your multimodal images during acquisition in order to plan your next steps. ZEN gives you multiple options. You can sit at your connected computer to start the new Direct Processing function for processing your Airyscan images during acquisition.

However, confocal imaging is only one part of the big picture, and you may need data from additional imaging modalities to complement the view on your sample.

ZEN Connect can bring information from all your experiments together. Keep the context of your data by collecting all images of one experiment session in a single project in which you can combine overview and detailed high-resolution images, all perfectly aligned. Once you have created a project, you can always add and align content from any other imaging source, be it ZEISS, non-ZEISS or even sketches and analysis graphs. You will stay on top of things at all times – both during your experiments and months or years later.

Your ZEN Connect projects keep all associated datasets together. It’s never been easier to share results and co-work with others as a team. The powerful integrated 3Dxl Viewer, powered by arivis®, is optimized to render the large 3D and 4D image data you have acquired with your fast new LSM 900. You can create impressive renderings and movies for meetings and conferences. After all, a good picture can say more than a thousand words.

Section of a Thy1-YFP mouse brain

© Courtesy of R. Hill, Yale University, New Haven, CT, USA.

Section of a Thy1-YFP mouse brain
Section of a Thy1-YFP mouse brain - single neuron

Section of a Thy1-YFP mouse brain. Thy-1 (green) is involved in the communication of cells in the nervous system. Overview image (A) acquired on ZEISS Axio Scan.Z1. Inset shows enlarged ROIs imaged on ZEISS LSM with Airyscan (B) The neuronal network is clearly visible. The depth of the Z-stack is color-coded. (C) shows a single neuron.


The micrograph shows a Lilium auratum pollen grain, acquired with Airyscan 2 in Multiplex mode.

The micrograph shows a Lilium auratum pollen grain, acquired with Airyscan 2 in Multiplex mode.
© Courtesy of Jan Michels, Zoological Institute, Kiel University

click to enlarge

Nuclei of living HeLa Cells were labelled with 5’-610CP-Hoechst (Chem.Sci. 2019, 10, 1962 – 1970). The dye is added to the cell culture media in a defined concentration. The bleaching experiment (FRAP) confirms that the dye needs about 15 minutes to efficiently label DNA. The time series is recorded for 13.5 minutes with 1 frame per second; with the bleaching event in the labeled region after the first 10 frames.  © Courtesy of P. Lenart, MPI for Biophysical Chemistry, Göttingen, Germany

Downloads

ZEISS LSM 900 with Airyscan 2

Your Compact Confocal for Fast and Gentle Multiplex Imaging

29 Pages
Filesize: 8,109 kB

The Basic Principle of Airyscanning

22 Pages
Filesize: 1,473 kB

ZEISS LSM 9 Family with Airyscan 2

11 Pages
Filesize: 3,114 kB