Once fluorescent proteins began revolutionizing biological imaging, the confocal microscope cemented its place as the centrepiece of the core imaging lab. The multitude of sample types and applications that benefit from optical sectioning ensure the confocal is always in high demand and for a large variety of imaging needs. ​

As experimental demands grow, so progress in technology presents new opportunities and the drive for faster acquisition with higher resolution and better image quality continues to expand experimental possibilities.​

The Airyscan detector provided a huge step forward in confocal performance when it was launched in 2014 as it uniquely teamed super resolution with low noise and extremely gentle confocal acquisition. Subsequent developments have further extended this Airyscan capability and these advancements in speed, resolution and gentleness together are providing exciting opportunities for addressing new biological questions. ​

Your goal may be understanding the interplay of fluorescently tagged proteins in cell culture, expanding your biology to 3D models such as organoids, following fast signalling processes deep in the mouse brain or capturing large volumes of fixed samples. Each of these requirements benefits from lower noise and better image quality. The combination of gentle acquisition, super resolution and speed provided by the Airyscan detector delivers this quality for every single one of your samples and it is this diversity of application benefits that places the Airyscan at the centre of so many imaging explorations. ​

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The power of visualizing complete model organisms and tissues in 3D continues to transform our understanding of biology. This demand for contextual data in larger specimens, both live and fixed, has led to a significant increase in the use of samples such as zebrafish, Drosophila, organoids and cleared tissue for probing cellular and subcellular structure and function. Capturing meaningful 3D datasets in such samples is challenging using classical imaging approaches due to limitations in depth penetration and the resulting long acquisition times and significant phototoxic burden that can result. ​

By switching from classical imaging approaches to acquisition using a sheet of light, large volumes of data can be captured with the speed and gentleness necessary for these delicate specimens.​

Light sheet technology not only increases your image capture rate in larger living samples but also in fixed and cleared tissue and the resolution now made possible by advances in optics is generating impactful results. Making use of this powerful imaging modality for both live and fixed samples all in the same instrument really diversifies the applications and users that can benefit and ensures that the system is always used to full capacity.​

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The drive for reproducibility and statistical robustness has fueled an increase in automated, efficient microscopy workflows to reduce user bias and quickly increase the number of sample points for analysis. Automated microscopes and associated analysis routines complement other imaging solutions by providing a route to rapidly scale up experiments without needing to know anything about microscopy at all. ​

Historically, increasing throughput by using automation has meant a reduction in image quality, but advances in technology no longer engender this compromise. The ZEISS Celldiscoverer 7 has been designed from the ground up to provide optimal image quality for each and every user and sample. Optimization is automatic and with the implementation of analysis tools such as the ZEISS Bioapps, users have an easy route to generate reliable, quantifiable data without the need for extensive training.​

A further opportunity presented by automated imaging is implementation of analysis routines which automatically localize the events of interest and subsequently capture these at high resolution entirely automatically. This automatic acquisition of the multi-modal datasets is vital to glean as much information as possible without needing to spend hours locating the regions of interest manually. ​

Whether your goal is high content screening, capturing multiplex data, timelapse acquisition or multimodal imaging, combining automated acquisition with quick and easy analysis routines is key to reducing the time needed to reach your results. ​

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Lattice light sheet microscopy hit the headlines following the landmark paper in 2014 (1) and has since proved itself to be a powerful technique for the long-term study of a wide range of specimens. The unique combination of fast, volumetric imaging with extremely gentle acquisition is generating a richness of data that simply cannot be reached using any other imaging approach. This promises to completely revolutionize the field of live cell imaging, and homebuilt instruments have already generated numerous high impact publications. ​

However, the scope and usability of this approach for a very wide user base has been limited since this technology has only been available in an upright configuration and has required significant time and expertise to make the necessary alignments and optimizations. ​

This changed with the launch of the ZEISS Lattice Lightsheet 7 platform which provides lattice light sheet imaging but in a convenient, inverted configuration. Compatible with the same samples and sample preparation you would use when imaging with a confocal or a spinning disc, the Lattice Lightsheet 7 offers an easy way to move existing experiments to this new capability without any reconfiguration.​

The automatic alignments and easy workflows provided by the Lattice Lightsheet 7 mean that even inexperienced users can access this cutting-edge approach and capture 3D data over hours and days at a time. The accessibility of such powerful technology to non-expert users sets this approach to become the new standard in 3D long term timelapse imaging of dish mounted specimens. ​

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1) Science (Oct 2014); 346(6208):1257998. Bi-Chang Chen et al, Lattice light-sheet microscopy: imaging molecules to embryos at high spatiotemporal resolution. DOI:10.1126/science.1257998

Super resolution light microscopy has played an important role in extending our understanding of biological processes in many sample types and contexts. A variety of technologies exist with each providing different benefits in terms of sample types, resolutions and light requirements. For example, single molecule localisation microscopy (SMLM) is inherently slow but provides very high resolutions (<10nm); Stimulated emission depletion microscopy (STED) enables high resolution imaging but requires special staining protocols and high laser powers that are known to damage live specimens; and super resolution structured illumination microscopy (SR-SIM) provides easy and gentle 3D insights, which are ideal for live specimens but historically with resolutions only down to 120nm, which is insufficient for resolving many sub-organelle processes and interactions.​

Those using super resolution microscopy on a regular basis, particularly for live imaging, are very familiar with these trade-offs in terms of speed, resolution and gentleness and until recently the technology platform of choice for each experiment has been the system providing the best compromise of these three parameters. ​

With ZEISS Elyra 7, these compromises are no longer necessary. Your demanding super resolution requirements are met by a single platform offering both SMLM and lattice SR-SIM. Not only do you benefit from the stability and power you need for PALM and dSTORM experiments but also the new capability of SIM² which provides resolutions down to 60nm but with twice the speed and several times the gentleness of any other SIM approach. This is an exciting new era of super resolution capability!​

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The very latest in imaging instrumentation is providing many new and exciting opportunities for your Life Science research. This progress in acquisition approaches is powerful but it needs to be combined with a subsequent workflow of visualization, processing and analysis steps to achieve the quantitative insights that your research needs.​

The tsunami of data that imaging now generates causes big challenges for software platforms. Software instability or a lack of functionality or automatization can lead to processed data that provides less information than it could. Indeed, selecting the best solution for extracting the information you need from your imaging data can mean the difference between reaching answers to your scientific questions or not. ​

To complement the exciting developments in imaging technology, ZEISS also has a powerful range of solutions to support your big data analysis requirements to ensure that you can always extract the insights you need.​

Arivis Vision4D has been designed from the ground up to optimize the visualization and processing of even the largest datasets. Powerful 3D analysis routines and scalable workflows can be quickly and easily configured, ensuring that you always have the latest processing tools to get all the information you need from your precious imaging data.​

During this workshop you will have the opportunity to explore some of the latest developments in image handling, visualization and processing, and a chance to discuss common challenges that can be overcome with modern data management and processing solutions.

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