ZEISS Lightsheet Z.1 Microscope

This paper describes theoretical and practical aspects of sample preparation for light sheet fluorescence microscopy (LSFM). It presents general rules for sample handling and mounting, as well as guidelines with respect to the best preparative technique to use, taking into account sample type, structure and properties. Step-by-step protocols and recommended materials for ZEISS Lightsheet Z.1 samples are included. These protocols cover sample preparation ranging from micrometer-sized fluorescent beads to millimeter-sized insects, providing detailed information relating to preparation and observation techniques. Finally, this paper identifies the main artifacts and problems that can result from the preparation techniques.

This application note demonstrates a novel technique to formulate the optimum refractive index (RI) without compromising image quality or damaging the solvent-vulnerable Light Sheet Fluorescence Microscopy (LSFM) system. LSFM uses a thin plane of light (light sheet) to optically section transparent tissues or whole organisms that have been fluorescently labeled. This “RI matching” solution is based on the published 3DISCO, an organic-solvent based protocol, which requires no special equipment to provide a faster imaging solution compared to laser scanning microscopes.

Scientists must consider specimen integrity, fluorescent labelling and refractive indices (R.I.) when working with clearing applications and adapting the clearing procedures to their specimens. It is crucial to check the R.I. of clearing solutions to minimize mismatches and resulting optical errors, such as spherical aberrations, and to increase the image quality and depth penetration, before and during the clearing procedure. This technology note describes a simple method to check the R.I. for clearing and imaging solution necessary for successful clearing imaging with Lightsheet Z.1.

Light sheet fluorescence microscopy has a wide range of applications including use in developmental biology, systems biology, organogenesis and cell dynamics, 3D cell culture, plants, imaging of marine animals as well as structural imaging of large, fixed specimens and imaging of optically cleared specimens. The Zeiss Lightsheet Z.1 allows you to observe model organisms, tissues and cells as they develop over days with virtually no phototoxicity or bleaching. This product note describes the benefits of Zeiss’ Lightsheet Z.1; the first light sheet microscope designed to image fluorescently labeled living samples.

The ZEISS Lightsheet Z.1 imaging system is capable of delivering optical sections of large samples, with virtually no phototoxicity or bleaching and with high temporal resolution. Light Sheet Fluorescence Microscopy has advantages for imaging live as well as cleared samples, with the Lightsheet Z.1 as equally capable in clearing applications. This document describes the necessary steps to change between water immersion and clearing immersion imaging options.

In the spring of 2013 the Harvard Center for Biological Imaging (HCBI) was able to demo, and eventually acquire, Lightsheet Z.1 microscope from ZEISS. The HCBI have now performed a number of long term embryonic development imaging experiments including imaging of zebrafish, fly, worm, cricket, and salamander embryos. Throughout these experiments, ZEISS Lightsheet Z.1 was employed to acquire both fast dynamics and long-term changes during development; this application note evaluates the use of ZEISS’ Lightsheet Z.1 in such experiments.

This white paper describes the theoretical and practical aspects of sample preparation for Light Sheet Fluorescence Microscopy. General rules for sample handling, as well as guidelines with respect to the best preparative technique to use are presented. Step-by-step protocols, covering sample preparation ranging from micrometer-sized fluorescent beads to millimeter-sized insects, are also included.

This technology note demonstrates the integration of daylight illumination into time lapse experiments.

This application note presents an application of the commercially available high-resolution light sheet based microscope, Lightsheet Z.1 from ZEISS, to the investigation of large cultured organotypic tissues, so called cellular spheroids.

Lightsheet Z.1 allows you to record the development of large, living samples or 3D tissue cultures and gently image them to deliver exceptionally high information content. In this video see how easy it is to prepare samples for imaging with Lightsheet Z.1 - the MultiView Light Sheet Fluorescence Microscopy system by ZEISS.

Hear how the Lightsheet Z.1 Multiview light sheet fluorescence microscope from ZEISS enables you to image large living samples in physiologically relevant conditions for long periods of time, with virtually no phototoxicity or bleaching. The system also allows for imaging of large cleared specimens, with exceptional light efficiency and speed.

Hear how ZEISS has worked together with arivis AG to provide a software platform for the Lightsheet Z.1, which can handle the large datasets produced by 3D imaging of large samples, enabling analysis of tiled stacks and large time dependent series. An example data set of a mouse hippocampus is presented.

In this video Scott Oleynch, Marketing Manager for Zeiss Microscopy, tells SelectScience how the new Lightsheet Z.1 microscope system enables scientists to perform fluorescence imaging on large, living samples, with virtually no phototoxicity or bleaching and with high temporal resolution. Scott discusses some of the unique features of the system and some of the applications for imaging model organisms such as zebrafish, c. elegans and drosophila. Interview filmed by SelectScience at AACR 2013.

Watch this video to learn how the innovative Lightsheet Z.1 from Carl Zeiss Microscopy enables you to perform 3D fluorescence imaging on large, living samples for prolonged periods of time, with virtually no phototoxicity or bleaching and with high temporal resolution. The unique Multiview Light Sheet Fluorescence Microscope allows you to record the development of large, living samples and gently image them to deliver exceptionally high information content. Watch this video to see examples of how the new Lightsheet Z.1 gives you the tools you need to work at unprecedented speed and acquire images of your whole sample volume at sub-cellular resolution in a fraction of the time it takes using other techniques.

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SelectScience® spoke to Dr Peter O'Toole, Head of Imaging and Cytometry, Department of Biology at the University of York, about the technology serving his department

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