Image with 4 – 8× more signal-to-noise ratio and with superresolution at highest frame rates.
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ZEISS LSM 900 with Airyscan 2

Your Compact Confocal for Gentle Multiplex Imaging and Smart Analysis

To pursue your scientific questions, you want only the best data quality. In microscopy, this translates into the best contrast and resolution while maintaining minimum light exposure. LSM 900, your compact confocal microscope, provides this with components optimized to deliver the best imaging results.

  • Get high-end confocal imaging in a small footprint.
  • Improve any confocal experiment with LSM Plus.
  • Push super-resolution and speed with Airyscan 2.
  • Increase your productivity with AI Sample Finder.
Live imaging with LSM Plus: Plasma membrane localized expression of Wnt3-EGFP in the developing optic tectum of 4-day-old zebrafish.
Courtesy of C. Teh, Centre for Bioimaging Sciences, Singapore
Courtesy of C. Teh, Centre for Bioimaging Sciences, Singapore

Live imaging with LSM Plus: Plasma membrane localized expression of Wnt3-EGFP in the developing optic tectum of 4-day-old zebrafish.

A Unique Confocal Experience in a Small Footprint

LSM 900 is packed with solutions for producing the best quality in confocal live cell imaging – with each component optimized for the highest sensitivity and contrast. LSM Plus lets you easily optimize the results of your multi-color and live cell experiments. All these high-end features fit in a small footprint and come with reduced complexity, so you’ll save valuable lab space and minimize the time required for user training.

Spermatogonia in Drosophila testis. Multi-color label with asterless (magenta), acetylated tubulin (cyan), and Hoechst 33258 (yellow). Imaged with ZEISS Airyscan 2 followed by Joint Deconvolution.
Courtesy of S. Song, Prof. Liou Yih-Cherng's lab, Singapore
Courtesy of S. Song, Prof. Liou Yih-Cherng's lab, Singapore

Spermatogonia in Drosophila testis. Multi-color label with asterless (magenta), acetylated tubulin (cyan), and Hoechst 33258 (yellow). Imaged with ZEISS Airyscan 2 followed by Joint Deconvolution.

Get Better Data – Faster

Airyscan 2 allows you to do more than any conventional LSM detector. Each of its 32 detector elements collects additional information, while all of them together gather even more light, yielding super-resolution quantitative results. By adding structural information with Joint Deconvolution (jDCV), you can push resolution even further. Or use the Multiplex modes to collect more information in less time.

ZEN Connect: From acquiring an overview image, to defining ROIs, and when changing between different imaging systems: You save time and always stay on top of things.

ZEN Connect: From acquiring an overview image, to defining ROIs, and when changing between different imaging systems: You save time and always stay on top of things.

Increase Your Productivity

ZEN microscopy software puts a wealth of helpers at your command to achieve reproducible results in the shortest possible time. AI Sample Finder helps you quickly find regions of interest, leaving more time for experiments. Smart Setup supports you in applying best imaging settings for your fluorescent labels. 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.

A Streamlined Light Path

Surprising Flexibility with up to Three Confocal Detectors

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 part of the light. You can define up to three detectors (multialkali, GaAsP or Airyscan 2).

  • Enjoy outstanding image quality and a superb signal-to-noise ratio.
  • Achieve faster scan speeds while preserving image quality.
  • Avoid photobleaching and phototoxicity by using low laser powers.
  • Detect faint signals in low expressing cells.
  • Use up to three spectral confocal channels simultaneously.
RPE1 cells transfected with H2B-GFP plasmid. Maximum intensity projection of 117 Z-planes. Without LSM Plus.
RPE1 cells transfected with H2B-GFP plasmid. Maximum intensity projection of 117 Z-planes. With LSM Plus.

LSM Plus

Improving the Whole Confocal Experience

LSM Plus improves literally any confocal experiment with ease, independent of detection mode or emission range. Its linear Wiener filter deconvolution needs next to no interaction while still ensuring a reliable quantitative result. Just as in our time-tested Airyscan super-resolution processing, the underlying optical property information is adapted automatically based on objective lens, refractive index, and emission range.

Apply LSM Plus with no extra effort and benefit from:

  • Enhanced signal to noise at high acquisition speed and low laser power – particularly useful for live cell imaging with low expression levels
  • Improved resolution of your multi-color and spectral datasets
  • More spatial information and even greater resolution enhancement for bright samples that allow to close the pinhole of the LSM
  • Integrated workflows to combine the advantages of LSM Plus with Airyscan super-resolution imaging

Caption: RPE1 cells transfected with H2B-GFP plasmid. Maximum intensity projection of 117 Z-planes. Comparison of without (left) and with LSM Plus (right).
Courtesy of Tingsheng, Mitosis Lab, Singapore

 Schematic beam path of ZEISS Airyscan

Schematic beam path of ZEISS Airyscan

ZEISS Airyscan 2

(1) Mirror, (2) Variable Secondary Dichroic, (3) Airyscan optics, (4) Airyscan detector, (5) Airy disk

Airyscan 2

A Unique Combination of Super-resolution Imaging and High Sensitivity

Classic confocal laser scanning microscopes use point illumination to scan the sample sequentially. A pinhole spatially limits the extended Airy disk to block out-of-focus light from the detector. Closing the pinhole gives higher resolution, but at the price of detecting fewer photons.

Airyscan 2 is an area detector with 32 circularly arranged detection elements. Each of these acts as a small pinhole, contributing to super-resolution information, while the complete detector area collects more light than the standard confocal setting. This produces much greater light efficiency while capturing enhanced structural information.

Comparing the confocal image (left) with Airyscan SR (middle) and Airyscan Joint Deconvolution (right). HeLa cell, 4x expanded and labelled with acetylated alpha tubulin (green).
Courtesy of S. Zhang, Prof. Liou Yih-Cherng's lab, Singapore
Courtesy of S. Zhang, Prof. Liou Yih-Cherng's lab, Singapore

Comparing the confocal image (left) with Airyscan SR (middle) and Airyscan Joint Deconvolution (right). HeLa cell, 4x expanded and labelled with acetylated alpha tubulin (green).

Comparing the confocal image (left) with Airyscan SR (middle) and Airyscan Joint Deconvolution (right). HeLa cell, 4x expanded and labelled with acetylated alpha tubulin (green).

32 Views Mean More Information

Powerful Deconvolution with Airyscan jDCV

Each of the 32 Airyscan detector elements has a slightly different view on the sample, providing additional spatial information that makes Joint Deconvolution possible. This reduces the distance that can be resolved between two points even further – down to 90 nm. Your super-resolution experiments will benefit from an improved separation of single or multiple labels.

Airyscan 2 with Multiplex

Large Fields of View and Whole Sample Volumes in the Shortest Time

The Airyscan Multiplex modes use knowledge of 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 larger steps when sweeping the excitation laser over the field of view, improving acquisition speed. Capturing more spatial information in the pinhole plane allows final image reconstruction with better resolution than the acquisition sampling.

  • Up to four image lines in a single sweep
  • Rapid tiling of large areas
  • Efficient live cell imaging
  • Fast volumetric imaging

Multiplex Modes of ZEISS LSM 900

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

++

+++

++++

+++++

AI Sample Finder

Automated Sample Identification for Efficient Imaging

For sample placement, microscope parts such as the condenser arm often have to be moved manually. Focus adjustment and identification of the relevant areas on the sample carrier require additional manual steps. AI Sample Finder automates this sequence, eliminating time-consuming manual adjustments and reducing the time to image from minutes to just seconds.

  • Access all sample areas directly.
  • Reduce your time to experiment to just seconds.
  • Easily image only those regions containing sample.
  • Don’t miss potentially important areas.

Applications

ZEISS LSM 900 at Work

Cell Division of LLC-PK1 (Porcine Kidney)

To minimize photobleaching and damage to a live sample, it is useful to reduce acquisition time and to use minimal laser power. LSM Plus helps to improve the signal-to-noise ratio as well as the resolution of structures such as spindle fibers.

In this example, 100 Z-stacks were acquired with LSM 900 on Celldiscoverer 7 over 29 minutes. The images show a maximum intensity projection of 38 Z-planes. Cells expressing H2B-mCherry (red) and α-Tubulin-mEGFP (cyan).

Zebrafish Embryo (2 Days)

LSM Plus helps to improve signal-to-noise ratio when imaging large volumes to be rendered in 3D. Visualization of the vasculature (green) and red blood cells (magenta) by transgenic reporter expression, lateral view, anterior to the left.

A 300 µm Z-stack with 81 planes over three tiles was imaged with LSM Plus applied. The tiles were stitched and rendered in 3D with ZEN – powered by arvis®.

Sample courtesy of B. Schmid, DZNE Munich, Germany

 Mitochondrial Structures in Cos7 Cells
 Mitochondrial Structures in Cos7 CellsSample courtesy of Zhang Y, University of Science and Technology of China, China
Sample courtesy of Zhang Y, University of Science and Technology of China, China

Mitochondrial structures in Cos7 cells

Mitochondrial structures in Cos7 cells

Mitochondrial Structures in Cos7 Cells

Images were acquired with LSM 900 on ZEISS Celldiscoverer 7 using confocal GaAsP detectors (top row) and Airyscan 2 in HS mode (bottom row). Confocal images with LSM Plus (top, right) enhancing SNR and improving resolution of mitochondrial structures. Airyscan Joint Deconvolution (bottom, right) resolves the inner and outer membrane architecture even better compared to Airyscan HS (bottom left). Stained for mitochondrial outer membrane protein Tom20 (Green, Alexa Fluor-488) and mitochondrial inner membrane protein ATP5a (Magenta, Alexa Fluor-647).

  • Multiplex mode
  • Airyscan SR
  • Comparing the field of view you can image at superresolution in the same time using Multiplex mode.
    Comparing the field of view you can image at superresolution in the same time using Multiplex mode.
  • Comparing the field of view you can image at superresolution in the same time using Airyscan SR
    Comparing the field of view you can image at superresolution in the same time using Airyscan SR

Superresolution and Multiplex Mode

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).

Multiplex Mode

Drosophily embryo imaged with Multiplex Mode for LSM 900.
Drosophily embryo imaged with Multiplex Mode for LSM 900.

Drosophily embryo imaged with Multiplex Mode for LSM 900.

Drosophily embryo

Imaged with Multiplex Mode for LSM 900. Courtesy of J. Sellin, LIMES, Bonn, Germany

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.Image courtesy of Jan Michels, Zoological Institute, Kiel University
Image courtesy of Jan Michels, Zoological Institute, Kiel University

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

Lilium auratum pollen grain

Acquired with Airyscan 2 in Multiplex mode. Courtesy of Jan Michels, Zoological Institute, Kiel University

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

Zebrafish Embryo (Danio Rerio)

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.

A.C. Hocke, Charité, Berlin, Germany.

Human Lung Epithelial Cell Line

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

The acquisition seemlessly combines two imaging modes – the fluorescent channels were 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.

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.

Bring Information From all Your Experiments Together

Overview image. Section of a Thy1-YFP mouse brain on ZEISS Axio Scan.Z1.

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

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.

Neuronal network
Neuronal network. Inset shows enlarged ROIs imaged on ZEISS LSM with Airyscan

The neuronal network is clearly visible. The depth of the Z-stack is color-coded.

Single neuron
(C) shows a single neuron.

Section of a Thy1-YFP mouse brain - single neuron

  • Nuclei of Living HeLa Cells
  • Nuclei of Living HeLa Cells. Time Series
  • Nuclei of Living HeLa Cells. Time Series

Nuclei of Living HeLa Cells

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

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    • ZEISS LSM 900 with Airyscan 2

      Your Compact Confocal for Gentle Multiplex Imaging and Smart Analysis

      Pages: 34
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      Multiplex Mode for Fast and Gentle ConfocalSuperresolution in Large Volumes

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