Microscopy from Carl Zeiss

Cell Observer® SD Two Pioneering Technologies in Perfect Harmony.

Carl Zeiss is combining two high-performance technologies: the high-end Cell Observer® microscopy platform and the CSU-X1 spinning disc technology from Yokogawa. Each one leads the way in its own segment. Together they guarantee high-speed confocal microscopy, precision and outstanding flexibility. A system concept for the most demanding applications: Cell Observer® from Carl Zeiss The high-end Cell Observer® platform was developed for complex Live Cell Imaging applications. At the core of Cell Observer® is the inverted Axio Observer research microscope – expertly coupled with specially designed objectives and a modular incubation system. Further integrated into the platform are highly sensitive cameras and piezo focusing devices. All in all, this is a system that offers accuracy and reproducibility even during extremely rapid image acquisition, due to the precise control of all components by the AxioVision software.

The Confocal Microscope System for Live Cell Imaging With Minimal Phototoxicity

Speed and excellent light efficiency: Yokogawa CSU-X1 For many years, Yokogawa’s spinning disc technology has set the standard in high-speed Live Cell Microscopy. The innovative technology is based on a tandem-disc system containing a microlens array and a Nipkow disc. This guarantees maximum scanning speed and light efficiency, as well as outstanding reliability. Bleaching and phototoxicity are reduced to a minimum. Besides extremely high frame rates and a fast filter wheel, the new Yokogawa CSU-X1 scanhead also offers a Dual Camera option, making it the ideal choice for high-speed, confocal image acquisition.

Unique Spinning Disc Technology and Research Platform Combined in One System. A New Level of Performance for Live Cell Imaging. When carrying out pioneering research, powerful technology is absolutely essential. Cell Observer® SD combines maximum frame rates and flexible functionality in all its components to offer the user a powerful overall system.

A question of precision: full integration of components Cell Observer® is a modular system that can grow as your requirements expand. The different components are coordinated perfectly with one another and complement each other with precision during experiments. The modular system architecture offers the optimum conditions for integrating Yokogawa CSU-X1. Artifact-free image acquisition is guaranteed – even at extremely high frame rates – and the integration of all components ensures maximum protection of the sample during the observation of living cells. This is a system that will impress thanks to its simple and convenient operation. Two technologies combined: one point of contact The entire Cell Observer® SD system combines unique technologies from Carl Zeiss and Yokogawa, yet with this system everything comes from a single source. The configuration, service and support are all from one point of contact you can rely on.

Cell Observer® SD system

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For a perfect climate: modular incubation systems Optimum cultivation conditions for sensitive samples – the Carl Zeiss incubation concept is as unique and as variable as the requirements it serves. From compact stage-top incubators to the XL incubator with particularly impressive temperature stability, anything is possible. Environmental controls are provided via stacking modules which can be easily added to a new or existing microscope. Whether it’s a question of temperature, CO2 control or O2 concentration, with the AxioVision software you are in control of all incubation parameters at all times. This enables you to perform reproducible live cell experiments under precisely recorded conditions. In addition, complete safety is always guaranteed thanks to an inventive laser safety system – without restricting operating convenience in any way.

Incubator PM S1 offers maximum space during operation and allows dynamic temperature experiments

Impressive efficiency: Laser module and filter sets The sophisticated fluorescence technology of Cell Observer® SD will perform for all of your requirements, no matter how demanding. The excitation light is supplied by a laser module with up to four powerful solid-state lasers and efficient coupling, while the lines and intensity are controlled by an AOTF. With a range of single- and multi-bandpass filters and beamsplitters, the system configuration can be adapted to your needs and application. One system for many applications Fast, flexible and convenient – Cell Observer® SD is the ideal microscope platform for a wide range of applications. Its high speed makes it possible to observe extremely rapid processes such as membrane transport in eukaryotic cells. Fast Multichannel Imaging allows different vesicle populations to be observed simultaneously in 3D over time and then to be tracked automatically. Interactions between cell compartments can be resolved in detail. Thanks to its low phototoxicity, Cell Observer® SD is also ideally suited for sensitive processes. Long-term observations of cell division can be performed with high contrast without causing damage to cells. Fast signal transduction processes can be followed using reporter dyes or FRET indicators with high temporal resolution. Here, confocal image acquisition allows signals to be allocated clearly and the Dual Camera option allows emission ratios to be acquired without channels being temporally out of step. And all this with maximum precision, even when it comes to highly complex applications.

Mitosis in LLC-PK1 cells, tubulin-EGFP + Histon 2b-mCherry Mike Davidson, University of Florida, USA

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Cell Observer® from Carl Zeiss. Maximum Performance Can Be Seen in the Details. When milliseconds count, all components must interact perfectly. The high-end Cell Observer® research platform guarantees seamless integration of all components and precision in the acquisition of extremely rapid processes. Plus maximum flexibility – all image acquisition dimensions can be freely combined. Superior fluorescence: Axio Observer from Carl Zeiss The inverted Axio Observer research microscope from Carl Zeiss forms the basis of Cell Observer® SD. This microscope impresses with its high degree of stability and expandability. The apochromatically corrected beam path guarantees brilliant contrasting and homogeneous illumination, while the intelligent operating concept facilitates simple and convenient operation. The right objective for all applications Choosing the right objective is a crucial success factor in Live Cell Imaging, and Carl Zeiss has just the solution for every application: • Plan-Apochromat High numerical apertures – outstanding in terms of color correction, resolution, contrast and flatness of field. • α Plan-Apochromat 100x/1.46 Maximum resolution and magnification with excellent light efficiency. • LCI Plan-Neofluar Multi-immersion objective – the correction ring allows adjustment to various sample conditions and different temperatures. • C-Apochromat Water immersion objective – unsurpassed brilliance and minimal aberrations for samples in an aqueous medium. Different cover glass thicknesses can be corrected.

Modular and flexible expansion options: components for every application Cell Observer® is a system that grows as your research grows. It can be upgraded and retrofitted in line with your individual requirements. • Motorized stages For Multiposition Imaging with excellent reproducibility. Travel range, speed and precision can be adapted to your application. • Piezo focusing Enables maximum speed and precision when acquiring Z-stacks. Either the objective is moved or just the sample holder on the stage. • ApoTome Optical sections of ideal thickness, irrespective of laser lines and with the full range of objectives. Used in addition to Yokogawa CSU-X1, it is as a second method for generating optical sections on the same microscope. Sensitivity and speed: the cameras Experiments in cell biology place extraordinary demands on the camera. Whether you need high resolution or extremely fast frame rates – Cell Observer® SD integrates the right model for your particular application. • CCD cameras from the AxioCam family AxioCam MRm: for universal use; AxioCam HRm: for maximum resolution using microscanning; AxioCam HSm: for extremely high frame rates.

High-aperture and high-magnification objectives from the Carl Zeiss range, ideal for use with Cell Observer® SD

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Zeiss AxioCam MRm (right) and Photometrics QuantEM:512 (left) with corresponding camera adapter on the Yokogawa CSU-X1

To capture the crucial moment: maximum frame rates In Live Cell Microscopy it is essential to capture dynamic processes in their entirety. Cell Observer® therefore has a high-speed mode for achieving maximum frame rates. In this case, the data is streamed directly to a dedicated hard disc. Image acquisition triggers all microscope components, meaning that even the most rapid processes are fully resolved.

• Mark&Find Automatic capture from multiple positions. • MosaiX Tile-by-tile imaging of large specimens at high magnification. These tiles are then combined to form a large overview image. • Dual Camera module For the simultaneous operation of two cameras (e.g. for FRET imaging). This enables optimal use of the Yokogawa CSU-X1 Dual Camera option. • Physiology For functional imaging of reporter dyes (e.g. ion concentration) or FRET Imaging. Extensive analysis functions even online during acquisition.

Simple, intelligent and powerful: the AxioVision software During image acquisition all components of Cell Observer® must interact quickly and with high precision. AxioVision, the system software from Carl Zeiss, ensures perfect synchronization of the components. And for every application a suitable method of acquisition is available.

AxioVision offers a wide range of display and analysis options. Three-dimensional data sets can be displayed perfectly (Inside 4D) and measured in 3D (3D Measurement), while organelle movements can be tracked (Tracking) and measurement protocols automated (Automeasure). The ASSAYbuilder module makes it possible to analyze high-content experiments intuitively.

• Cameras from third-party manufacturers Compatible with the EM CCD cameras of Hamamatsu and Photometrics – for applications where each individual photon counts.

Fast MosaiX

Dual Camera

HDR Imaging

Widefield Multichannel Unmixing

Digital High Speed Recorder

Fast Acquisition

Imaging Plus

ASSAYbuilder

ELISPOT

Panorama

Mark&Find

ApoTome

3D Measure

TMA

SFM

MosaiX

Time Lapse

3D Deconvolution

QuantiFISH

Ratio

Tracking

Extended Focus

Z-Stack

2D Deconvolution

AutoMeasure

AutoMeasure Plus

Colocalization

Autofocus

Multichannel Fluorescence

Inside4D

Interactive Measurement

Online Measurement

Physiology

Image Acquisition

Image Processing

Image Analysis

VBA

Asset Archive

Commander

Documentation

Configuration

Modules relevant for Cell Observer® have been highlighted in orange

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Yokogawa CSU-X1. An Established Technology Which Now Impresses With Even More Power. Complex tasks demand extraordinary solutions. The unique technology of Yokogawa CSU-X1 guarantees speed and an excellent standard in Live Cell Microscopy. The simultaneous scanning of hundreds of points ensures high frame rates, while microlenses ensure maximum light efficiency. For extremely high frame rates: 1,000 points simultaneously The CSU-X1 confocal scanning unit manufactured by the Yokogawa Electric Corporation scans hundreds of points simultaneously. Considerably higher frame rates can be achieved compared to traditional point scanners. For the detection of fluorescence, EM CCD cameras with high quantum efficiency can be used and subsequently illumination intensity and phototoxicity are are minimized.

Time lapse Imaging of an endocytic vesicle fusing with a lysosome, time scale in seconds, HeLa cells, flotillin-2-EGFP + LysoTracker Elisa May and Daniela Hermann, University of Konstanz, Germany

A question of speed: the Nipkow disc These achievements are based on a unique technology: thousands of pinholes are arranged in a spiral on a rotating disc – the Nipkow disc. By illuminating and detecting through the disc, just as many points are imaged in parallel. By rotating the disc, the entire sample is gradually scanned. Once it rotates 30°, the camera‘s entire field of view is completely imaged once.

Widened laser beam Microlens array Dichroic mirror

Lens Pinhole array

Objective

Sample

Schematic diagram of the beam path in the Yokogawa CSU-X1

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Sample

CCD

Excellent light efficiency guaranteed: the tandem-disc system with microlenses Since a large proportion of the incident light would ordinarily be blocked in the area between the pinholes, Yokogawa CSU-X1 has been equipped with a special tandem-disc system: a second, mechanically coupled disc has been added in front of the Nipkow disc. This second disc contains microlenses which focus the incident excitation light precisely through the pinholes of the Nipkow disc and, therefore, ensures efficiency.

Yokogawa CSU-X1 with high-speed filter wheel (left) and Dual Camera option (right) – naturally both of these can also be combined

As flexible as the applications: different configuration levels There are a variety of applications which demand fast, confocal image acquisition. The CSU-X1 from Yokogawa has different configuration levels to best suit these various requirements: Rotation speed – Depending on the dynamics of the applications, it is possible to vary the rotation speed of the Nipkow disc. You can choose from a fixed speed of 1,800 rpm or adjustment between 1,500 and 5,000 rpm.

Versatile and fast Whether it’s a question of observing vesicle transport or signal transduction in living cells or whether 3D reconstructions of dynamic processes or ultra-fast 2D time lapse series are required: flexibility and speed make Yokogawa CSU-X1 the option for confocal image acquisition with the handling of a conventional widefield system. * purely mechanical switching time

Filter wheels – Optional high-speed filter wheels allow emission filters to be changed with a switching time of just 33 ms* between neighboring positions. Combined with an AOTF-based selection of excitation wavelengths, high-speed multichannel experiments can be realized. Dual Camera – The Dual Camera option enables the simultaneous acquisition of two different emission ranges using two cameras, ensuring that the two channels are temporally synchronized. As a result, e.g. colocalizations or ratio calculations are free of temporal inaccuracies.

3D rendering of a mouse stomach section, Sytox Green + Phalloidin-Alexa568 Mike Davidson, University of Florida, USA

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www.zeiss.de/CellObserverSD

60-2-0021/e – printed 10.08

BioSciences | Göttingen Location Phone: +49 551 5060 660 Telefax: +49 551 5060 464 E-Mail: [email protected]

Information subject to change. Printed on environmentally friendly paper bleached without cholorine.

Carl Zeiss MicroImaging GmbH 07740 Jena, Germany