Workshop 1

Modern microscopy techniques acquire an ever-increasing amount of multidimensional image data. 3D microscopy techniques, such as selective plane illumination microscopy (SPIM), easily generate multiple terabytes of raw image data in a single experiment that require tailored solutions for interactive or automated analysis.

The ACQUIFER HIVE is the established solution for efficient and safe management and storage of large image data without USB drives or network copy. The HIVE allows fast access to acquired data for swift processing and flexible analysis workflows. It is designed as a modular platform that is configured for your specific applications and will grow with your needs.

In this workshop, we will introduce the concepts of HIVE workflows, and we will present new solutions to expand existing HIVE installations to serve a higher user base and larger projects. Multi-GPU and Multi-HIVE solutions are outlined. Besides scaling in processing, concepts about data safety with large storage pools are discussed.

As part of the workshop, we offer RDP access on the centralized multi user system HIVE for testing. We encourage participants to bring their own laptops.

Please refer to our website at www.acquifer.de or call us at +49 (6221) 435 2000 for more detailed information on ACQUIFER products.

One of the core facilities’ duties is to provide end-users, usually researchers in life sciences, a fleet of microscopes at a level of performance compatible with their experiments. This is not an easy task because the performance of such devices tends to fluctuate or deteriorate over time for many reasons: misusing, aging, environment fluctuations, etc. This is especially true for high end imaging systems such as confocal or super resolution microscopes. Having access to a unique, reliable, and easy-to-use device to ensure microscopes’ performance would certainly make easier this tedious task.

To get quantitative and reproducible data, assessing the performances of fluorescence microscopes is a prerequisite before any imaging campaign. For example, system co-registration accuracy should be evaluated before any co-localization study; System field uniformity and intensity response before any study where intensity in the image matters; Spatial resolution before any study aiming at counting objects close to each other, etc.

Three years after our last presence at ELMI, we are happy to come back and present during this workshop the novelties we have developed in the meantime: improved hardware products, new analyses from 3D patterns, new analyses related to non-Argolight products, improvements in user interface and user experience, etc. Above all, the presentation aims to show how the quality control of fluorescence microscopes can be standardized with Argolight software solutions, and how the generated quality control data can be managed and centralized for later reporting.

In this workshop we will demonstrate Bruker super-resolution and light-sheet microscopy solutions (Luxendo SPIM family).
The SPIM demonstration will cover different sample types and potential applications ranging from life imaging of delicate live specimen such as organoids and embryos (mouse, zebrafish etc. ) to whole in-toto imaging of large cleared samples such es entire adult mice. We will be showing our 3D Super-resolution system, the VXL with patented 3D technology suitable for STORM and PALM, along with an integrated
microfluidics system for highly multiplexed, DNA-PAINT research.

Workshop 1/4 : TruLive3D SPIM – light-sheet microscopy for live samples
Workshop 2/5 : VXL – 3D super-resolution microscopy
Workshop 3/6 : LCS SPIM – light-sheet microscopy for cleared samples

Image:
Top, left image: live zebrafish embryo H2A::GFP recorded on Luxendo MuVi SPIM LS. Top, right image: Mitochondria stained for TOM20 with Alexa 647 and imaged with STORM. Bottom, left image: cleared adult mouse with GFP labeled nerves and recorded on Luxendo LCS SPIM. Bottom, right: Cleared YFP expressing transgenic mouse brain recorded on Luxendo MuVi SPIM CS

The launch of the ZEISS Lattice Lightsheet 7 inverted platform in 2020 was an important milestone for lattice lightsheet microscopy. This platform was designed for use by the non-expert so no alignment or technical know-how is needed to capture data. This has led to wide adoption of the Lattice Lightsheet 7 and it has now emerged as one of the key solutions for long term live imaging in the core facility. Further expansion of the Lattice Lightsheet 7 platform to more capability both in acquisition and analysis to assist in scaling experiments to higher throughput now further extend the imaging possibilities.

This workshop will focus on lattice lightsheet technology and will introduce the new capabilities that the ZEISS Lattice Lightsheet 7 offers. You will see the full workflow in action from capturing the data through to segmentation and quantitative analysis of the resulting datasets using arivis Vision 4D.

Fast and highly resolved volumetric microscopy is essential to examine various biological aspects in their complexity, ranging from whole animal imaging, to live phenomena, to thick samples to image at the different depth. During this workshop we demonstrate how X-light V3 spinning disk and DeepSIM add-on can be combined to work with deep samples of thickness comparable to those normally used in CF microscopy. This means that more meaningful data can be obtained from native heterogeneous complex (over 100µm thick) samples using routine preparation protocols.
#deepimaging #clearedtissues #spinningdisk #SIM

Characterising, comparing, and ultimately understanding, differences in dynamic live cell behaviour is critical in many fields, from cancer research to regenerative medicine.

In this workshop, we explore new ways to characterise cell behaviour, enabled by the Livecyte Kinetic Cytometer system. We look at how simple label-free assays, in 96-well plates, can be used to independently measure cell growth and cell proliferation, automatically characterise both random and sheet cell migration and identify other phenotypic changes. We also show Livecyte’s correlative fluorescence capabilities, and how these measurements unlock additional powerful new insights.

High-resolution microscopy of cells requires imaging chambers that are specifically tailored to your application.

Are you interested in learning about…
• … easy to handle all-in-one solutions for live cell imaging?
• … the advantages of using channel slides for immunofluorescence stainings?
• … solutions for 3D cell culture and spheroid imaging?

Join our workshop and find the answers to these questions!

First, we will introduce a variety of ibidi chambers with different geometries and surfaces and help you to choose the products that are best suited for your assay. This will be followed by a hands-on-training where you get the chance to test a selection of our labware for 2D and 3D cell culture and imaging.

Experience Mica in action and learn how its sample-centric operation allows straightforward access to imaging for all users .
See how MICA dramatically reduces the time-to-image and training time for non-imaging specialists.
Learn how our new FluoSync technology detects four fluorophores simultaneously, in both widefield and confocal!
Discover how the whole workflow from setting up an imaging experiment to the AI powered extraction of data has been radically simplified.
See all this working directly on samples and specimens you see every day – from multi-well plate assays to large tissue sections to model organisms – all on Mica!

Cutrale, F., Trivedi, V., Trinh, L. et al. Hyperspectral phasor analysis enables multiplexed 5D in vivo imaging. Nat Methods 14, 149–152 (2017). https://doi.org/10.1038/nmeth.4134
·Hsiao Chiang, Daniel Koo, Masahiro Kitano et al. HyU: Hybrid Unmixing for longitudinal in vivo imaging of low signal to noise fluorescence, 12 January 2022, PREPRINT (Version 1) available at Research Square [https://doi.org/10.21203/rs.3.rs-1073331/v1]

White paper: https://webcdn.leica-microsystems.com/fileadmin/global/products/Supercharge-the-way-you-work/White_paper_FluoSync_MC-0003306.pdf

Visualizing the three-dimensional architecture of complex organisms while analyzing biological processes in a whole-organ fashion are becoming the new standard in research. To provide a complete, smooth, and hassle-free 3D imaging workflow, Miltenyi Biotec covers this entire process offering solutions for sample staining, clearing and imaging. In this workshop, we will guide you through the most important steps of sample preparation, including a live demonstration, and will demonstrate how easily high quality 3D images of large samples can be acquired with our cutting-edge light sheet system, the UltraMicroscope BlazeTM.

With AX’s new capabilities and easily managed lifetime experiment combination, we add an extra dimension to confocal microscopy.
In this workshop, you will find out how one can acquire and visualize FLIM data with the help of NIS-Elements. Discover a quick and intuitive basic analysis package directly included in NIS-Elements: Large image, Z stack, automated complex acquisitions, and basic analysis. Easy export of FLIM image to Symphotime within one click for more in-depth is at reach.
There is much more to discover, so join us and learn how AX/FLIM provides seamlessly integrated lifetime acquisition with unprecedented resolution.

Studying spatial protein organization or protein copy number, clustering and mobility at the molecular level is extremely important from both a diagnostic and treatment perspective. Looking at cellular structures beyond the diffraction limit with a resolution of 15-20 nm with robust quantification has been enabled with Single-molecule localization microscopy (SMLM) like STORM or PALM. ONI has created the world’s first desktop super-resolution microscope, the Nanoimager, which is designed to operate on a standard lab bench and has a footprint smaller than a piece of A4 paper, with the most intuitive and simple software interface, making it also accessible to researchers not specialized in high end microscopy techniques with one major goal: make it accessible and simple to use.

It is really important to enable users to extract quantitative information from their localization-based images. We have done this by designing a cloud-based analysis software called CODI (COllaborative DIscovery). CODI helps users analyze their single-molecule data by extracting quantitative information on protein clustering, cluster size and area, cluster density, and so much more. It enables all researchers to upload their data, share or collaborate with other scientists, whilst gaining meaningful insights about their data.

During this workshop we will perform a two-color STORM on mitochondrial samples (TOM20 and HSP60) and reveal the sub-mitochondrial compartmental organization with 20 nm resolution. The aim is to introduce SMLM imaging as a powerful tool for rapid and multiplexed characterization of cellular structures along with a visual and quantification analysis platform CODI developed by us. We will follow up the workshop with an open discussion on the wide-ranging applicability of the Nanoimager in different biological applications encompassing the field of immunology, neurobiology, cell biology and cancer.

Bring your own sample to the conference to image it with Prospective Instruments MPX series Two-Photon Microscope. Contact us if you would like to take this opportunity.
Of course, we will also bring a selection of samples to image.

FLUCS photomanipulation is a non-invasive, all-optical method based on thermoviscous flow that allows the microscopist to induce precise microscopic flow fields inside living cells or microfluidic chambers.

During the workshop, we present RAPP’s FLUCS add-on optical module that is compatible with all standard inverted microscopes, we explain the working principle of FLUCS technology, we discuss a variety of recent applications of this novel technology in the life sciences, and we provide guidance, to optimally exploit the technology for your experiment.

www.rapp-opto.com/products/photomanipulation-systems/flucs-micro-flow-photomanipulation/

During these workshops you will discover, through a step-by-step demo, the recently added functions of the Thermo Scientific™ Amira™ Software’s new Xplore5D extension, which offers data conversion and compression with immediate visual feedback, allowing you to quickly interact with and communicate on large 3–5D data.

Tissue Cytometry permits to determine the in-situ phenotype of cells as well as histological entities, like glands, vessels or tumor foci. Applications include but are not limited to the exploration of the cellular/tumor microenvironment and/or the spatial organization of cellular subpopulations, assessment of different bone structures, quantification of blood vessels and neovascularization as well as analysis of samples in multiplexing or multispectral mode.
Earlier attempts to analyse single cells in tissue have mostly been subject to visual estimation, or – at best – to manual counting for decades. Hence, experts usually had the choice of the “least of evils” between guessing and endless (manual) counting. In (tumor) immunology, infiltrating inflammatory cells need to be phenotypically characterized on a quantitative basis. To better understand the function of inflammatory cells in tumor development, type and number of inflammatory cells and their proximity to glandular/tumor structures have to be analyzed in-situ and correlated with disease state. Using TissueFAXS™ Cytometry the time-consuming and error-prone human evaluation of stained histological sections can be approached with an observer-independent and reproducible technology platform, offering a high degree of automation, paired with user interaction at relevant points of the analytical workflow. TissueFAXS Cytometry is the “one-stop shop” for all kinds of microscopic analysis of tissue sections, organoids and adherent cell cultures.

Biological systems are complex and many environmental parameters affect their behavior. The chemical conditions of the sample volume, including pH and salt concentration, are often well considered when designing experiments in high and super-resolution microscopy. However, control over the physical conditions such as temperature or the amount of light deposited in the sample volume are often not well characterized and might differ from setup to setup. The imprecisions mainly arise due to the lack of appropriate sensor that can be placed close to or directly in the sample volume. Integrated systems, which are in direct contact with the sample, can help to improve reproducibility in biophysical studies of living cells, proteins or DNA and make interpretation of the acquired data easier.

Measuring and controlling the temperature of small sample volumes, as is often the case in high and super resolution microscopy, is technological challenging. In the first part of this talk, we will review state of the art approaches for temperature management in light microscopy, analyze their strengths and weaknesses and talk about our solution, VAHEAT. We will highlight the importance of controlling the temperature in biophysical studies based on recent research covering the fields of single molecules studies, live cell imaging and colloid chemistry.

In the second part of the talk, we will focus on illumination conditions specifically in total internal reflection (TIR) microscopy. After analyzing the limits of current technologies, we take the opportunity to pre-launch QuScite – our device for TIRF microscopy on a chip. This waveguide TIR system relies on photonic integrated circuitry (PIC) and allows to separate the excitation from detection path, which opens up completely new pathways for studying single molecules and processes in cell membranes. QuScite’s unprecedented field of view in the range of square millimeters combined with its ease of use even in complex workflows should allow everyone, on any microscope to perform high sensitivity TIRF measurements under reproducible conditions.

Note: This presentation will be interactive. You will have the opportunity to see and test demo systems on site.