Microscopy from Carl Zeiss

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Daniel Griffith
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Microscopy from Carl Zeiss LSM BiG Take the BiG Step Towards Universal Detection The BiG module s two GaAsP photodetectors deliver exceptional sensitivity, bringing you multiple detection modes and

Fit the BiG module to your existing system and get set to expand your potential in imaging, multiphoton imaging, photon counting, or FCS applications.

1 Microscopy from Carl Zeiss LSM BiG Take the BiG Step Towards Universal Detection The BiG module s two GaAsP photodetectors deliver exceptional sensitivity, bringing you multiple detection modes and twice the quantum efficiency of conventional detectors. The LSM binary GaAsP module BiG for short is a universal extension that expands your imaging productivity in a major way. Fit the BiG module to your existing system and get set to expand your potential in imaging, multiphoton imaging, photon counting, or FCS applications. The BiG module adapts to LSM 710, LSM 780, LSM 710 NLO and LSM 780 NLO systems. Until now, you ve had to choose between detectors will it be high dynamics, high sensitivity, or counting ability? While any one of these detectors may have reached an impressive level of performance, it comes at the expense of versatility. In addition, you would pay the less than optimal price for an upgrade for example, APDs for the most sensitive imaging. Now, LSM BiG s GaAsP technology introduces the potential for truly universal detection with the highest sensitivity, great dynamics, and precise counting ability all together in a compact module which can be fitted as an upgrade on most LSM 7 systems, adding two high performing detection channels to your existing ones. That s why this LSM BiG module more than lives up to its name. Universal Module for High Performance Detection. With LSM 710, LSM 780, LSM 710 NLO and LSM 780 NLO.

Fit the LSM BiG to Extend Confocal Detection.

You can use the additional channels in three modes: integration, photon counting, and FCS.

the population density and mobility of single molecules in the confocal volume.

You want to know how single molecules or defined molecule ensembles behave in your reaction tube or your cellular compartment?

2 Fit the LSM BiG to Extend Confocal Detection... Attaching the LSM BiG module to the direct coupling (DC) port of the LSM 710 scanning module lets you tap into two highly sensitive GaAsP detection channels. You can use the additional channels in three modes: integration, photon counting, and FCS. While integration and photon counting will extend the sensitivity for improved imaging of weak fluorescence signals up to a factor of two, the FCS mode lets you analyze the population density and mobility of single molecules in the confocal volume. As a bonus, the GaAsP detectors react extremely fast and are able to trace over a wide dynamic range. LSM BiG on direct coupling port (DC) of a LSM 710 or LSM 780. You want to know how single molecules or defined molecule ensembles behave in your reaction tube or your cellular compartment? You want to learn about their dynamic behavior and their preferred interaction partners? Then FCS will be an ideal choice. It measures precisely with highest sensitivity and quickly within seconds. Based on the confocal detection principle, the diffusion speed and concentration of a fluorescent labelled molecule are easily accessed in a single measurement by the correlation function. The GaAsP detectors allow integration and counting detection modes. Areas with the best count rate or interesting regions can be chosen in the image. 2

Both extend the sensitivity to improve the imaging of weak fluorescence signals, even when they are located very deep in the sample.

3 ... Extend Non-Descanned Detection too! You can also use the LSM BiG module as a non-descanned detector (NDD) on the LSM 710 NLO and LSM 780 NLO systems. This provides you with two highly sensitive GaAsP detection channels for multiphoton imaging to use in integration and photon counting mode. Both extend the sensitivity to improve the imaging of weak fluorescence signals, even when they are located very deep in the sample. You can use two BiG NDD modules at the same time for reflected and transmitted light detection. And the LSM BiG can also be combined with the conventional NDD modules of our NLO systems. Pulsed, ultrafast infrared lasers of NLO systems excite fluorescent dyes at the focus only the only place where photon density is high enough to produce fluorescence. With low absorption and next to no scattering loss, the infrared excitation light penetrates much deeper into the sample. Thanks to the small excitation volume, images of subcellular resolution are obtained. Since it s only at the focus where the dyes are excited (and where the dye can bleach), preservation of your fluorescence-labelled specimen is unmatched. LSM BiG as NDD detector on the sideport of an Axio Observer. Synapses and selected neurons in Drosophila melanogaster, labelled with Alexa anti-xfps. Multiphoton imaging with the LSM BiG in NDD position, projection of a Z-Stack. 5

Think BiG Upgrade Your LSM 710 and LSM 710 NLO System We offer various ways to upgrade your system with an LSM BiG module, depending on the scan module and light microscope of your own set-up.

4 Think BiG Upgrade Your LSM 710 and LSM 710 NLO System We offer various ways to upgrade your system with an LSM BiG module, depending on the scan module and light microscope of your own set-up. The direct coupling port can be retrofitted to the scanning module in the factory if not already available, enabling the LSM BiG upgrade itself to be carried out on site. For spectral confocal or FCS use, LSM 710 and LSM 780 with Axio Imager, Axio Examiner, and Axio Observer offer the following possibilities: LSM BiG can be mounted to the DC port of the scanning module Allows internal confocal and FCS detection LSM BiG channels can be combined with internal spectral detection No support column is required For multiphoton use, LSM 710 NLO and LSM 780 NLO with Axio Imager, Axio Examiner and Axio Observer offer the following possibilities: LSM BiG can be mounted to the reflected light NDD ports of Axio Imager, Axio Examiner and Axio Observer LSM BiG can be mounted to the transmitted light NDD ports of Axio Examiner and Axio Observer Allows multiphoton Imaging with highest sensitivity LSM BiG channels can be combined with 2 more standard NDD channels Support stands are available for Examiner and Imager if more than one module is used LSM BiG can also be mounted to the DC port in parallel to one NDD port unit For internal confocal and FCS detection, or NLO imaging with adjustable pinhole Two additional channels can be combined with the internal spectral detection No support columns required for DC port mounting Axio Imager, LSM on tube: BiG NDD, BiG DC Axio Observer, LSM rearport: BiG NDD, BiG DC Axio Observer, BiG DC 3

Beampath and Filterblocks for Your Applications Depending on the coupling

To guarantee best results, the filters are exchangeable by the user and

A choice of filtercubes for DC and NDD use is available.

filterwheel allows flexible settings of the detection wavelength range for

Using the LSM BiG as a NDD, standard NDD filtersets and beamsplitters allow

Filters for spectral confocal / FCS use Filterset 1 violet/green (BP 380-430

CFP/YFP (BP 455-500 / BP 525-570) Filterset 4 green/red (BP 500-550 / BP

non-descanned use Filterset 6 INDO (BP 380-430 / BP 465-515) Filterset 1

5 Beampath and Filterblocks for Your Applications Depending on the coupling position, the LSM BiG module sees different signals and wavelengths. To guarantee best results, the filters are exchangeable by the user and mounted in a standard P&C filtercube. A choice of filtercubes for DC and NDD use is available. In the LSM 710 and LSM 780 scanning modules, a secondary dichroics filterwheel allows flexible settings of the detection wavelength range for the LSM BiG. Using the LSM BiG as a NDD, standard NDD filtersets and beamsplitters allow the detection of any color excited by the multiphoton laser. Filters for spectral confocal / FCS use Filterset 1 violet/green (BP / BP ) Filterset 2 blue/green (BP / BP ) Filterset 3 CFP/YFP (BP / BP ) Filterset 4 green/red (BP / BP ) Filterset 5 yellow/red (BP / LP 650) Filters for non-descanned use Filterset 6 INDO (BP / BP ) Filterset 1 blue/green (SP 485 / BP ) Filterset 7 CFP/YFP (BP / BP ) Filterset 2 green/red (BP / BP ) LSM sideport: Axio Examiner, LSM on tube: BiG NDD Patents: Beampath and exchangeable filtercube in the LSM BiG. 4

Highly modular extension for universal detection Highly sensitive multicolor imaging RICS, FCS cross-correlation 3D in-depth

image yeast cells on a confocal yet. With the GaAsP detector we can do this now.

The GaAsP detector allows to speed up acquisition by a factor of two, with the same superb image quality as usually achieved by

Comments of LSM 780 and LSM BiG beta testers from labs in France, the United Kingdom, and Germany.

6 Highly modular extension for universal detection Highly sensitive multicolor imaging RICS, FCS cross-correlation 3D in-depth imaging Multiphoton imaging with adjustable pinhole Non-descanned detection with highest sensitivity We haven t been able to image yeast cells on a confocal yet. With the GaAsP detector we can do this now. also, very low expressing fluorescent proteins in cultured cells can be imaged in a new quality class. The GaAsP detector allows to speed up acquisition by a factor of two, with the same superb image quality as usually achieved by averaging twice on the PMTs. It s a great system, we really like the image quality. Comments of LSM 780 and LSM BiG beta testers from labs in France, the United Kingdom, and Germany. Carl Zeiss MicroImaging GmbH Jena, Germany BioSciences Jena Location Phone : Telefax : micro@zeiss.de Information subject to change. Printed on environmentally friendly paper bleached without chlorine /e printed 09.09

FLIM/FFS Upgrade Kits for Zeiss LSM systems

TECHNICAL NOTE FLIM/FFS Upgrade Kits for Zeiss LSM systems Shih-Chu Liao, Yuansheng Sun, Ulas Coskun ISS, Inc. 1 Introduction The FLIM and FFS Upgrade Package for the Zeiss confocal microscopes (LSM780,