Seeing is Believing: Imaging the Molecular Processes of Life

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Financial Assistance

Registration Fee Waivers

All academic and student registrants are invited to apply for a registration fee waiver, provided by the EMBL Advanced Training Centre Corporate Partnership Programme and EMBO. The registration fee waiver covers the registration sum that you have paid to attend the meeting. Conference participants are not required to pre-pay the registration fee to be selected for a fee waiver for a virtual meeting. If you have already paid the registration fee and are awarded a fee waiver, it will be reimbursed after the meeting. Course participants are required to pay the course fee in advance, which will then be reimbursed after the recipient has attended the course.

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For participants and speakers with childcare responsibilities there is the possibility to apply for a grant, provided by the EMBL Advanced Training Centre Corporate Partnership Programme and EMBO, to offset childcare costs incurred when participating at a virtual event. Eligible costs include fees for a babysitter or childcare facility or travel costs for a care giver. Please note that priority will be given to early stage researchers. Costs will be reimbursed after the meeting only once a reimbursement form and original receipts have been received. Attendance at the event is required in order to be eligible to receive the reimbursement. In order to apply for this grant, you must be registered by the abstract submission deadline.

Application

Applications for financial assistance can be submitted via the submission portal* (for the submission of abstracts for conferences or the submission of motivation letters for courses) by completing the Financial Assistance Application Section (underneath the section for entering abstract/motivation letter information). The link to the portal can be found in the registration confirmation email that you will receive after registering for the conference or course.

For conferences, if you are not submitting an abstract, you can still apply for financial assistance in the submission portal. Read the instructions on how to apply for financial assistance.

Note that priority will be given to those submitting an abstract to present at the conference. In your application you will be asked to answer questions regarding your motivation for applying, and, for registration fee waivers, the reasons why your lab cannot fund your attendance and how your attendance will make a difference to your career. Application for financial support will not affect the outcome of your registration application.

*For some events, applications for Childcare Grants will still be done by email. Information about the grant will be sent out shortly after the abstract/motivation letter deadline. Please contact the event Conference Officer if you have any questions.

Selection

The scientific organisers will select the recipients of registration fee waivers during the abstract selection process for conferences and the participant selection process for courses. Results will be announced approximately 3 – 4 weeks before the event start date. Selection results do not impact your admission to the meeting. Registration fee waiver selection is based on your current work or study location, your motivation for applying, the reasons for needing financial support and the impact this event will have on your career. Childcare grants are allocated based on career stage, with priority given to early stage researchers.

Further details

Check out this list of external funding opportunities or get more information on attending the conference as an event reporter.

For further information about financial assistance please refer to the FAQ page.

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Additional information can be found in our Code of Conduct.

Health and well-being

It is important to stay healthy and move around, especially when you are attending an event virtually. We have put together a few coffee break stretches and yoga videos in the events Slack workspace for you to enjoy during the event.

How to ask questions

Questions during and after the talks can be asked in live streaming platform. The chair moderates the questions and shares them with the speaker. If time runs out or you think of a question later, you can use Slack to ask your questions in the dedicated session channels or via direct message.

Time zone

The programme is planned based on the Europe/Berlin time zone, unless otherwise stated. As many virtual participants are attending from around the world, we do our best to accommodate as many time zones as possible when creating the programme. Please take your time zone into consideration when planning your attendance.

Virtual event platforms

We are using a virtual event platform for this conference. More information about the platform will be shared ahead of the conference.

SPEAKERS:

Dr. Julia Roberti, Product Manager Advanced Confocal Imaging
Dr. Irmtraud Steinmetz, Application Manager Advanced Confocal Imaging
Dr. Julia Koenig, Product Manager EM Sample Preparation for Cryo-workflows

ABSTRACT:

Live imaging is transforming our understanding of biological systems, enabling direct observation and quantitative analysis of structure and processes in cells, tissues, and organisms. The STELLARIS confocal platform provides the tools to achieve excellent image quality, spatiotemporal resolution, and functional information in close-to-physiological conditions. In addition, correlative correlative light and electron microscopy allows to directly identify the right cell at the right time and put dynamic live cell data into the ultrastructural context.

In the first part of the session, we will introduce the Stellaris confocal platform. The combination of the new generation white light lasers, acousto-optical beamsplitter (AOBS), and the Power HyD detector family, enables optimal tuning of both excitation and detection to achieve the most efficient signal acquisition in gentle conditions. Our new, proprietary approach to photon counting, Power Counting, significantly improves image contrast and delivers quantitative results. STELLARIS also includes Dynamic Signal Enhancement (DSE) and the new AI Feature AiviaMotion, a powerful digital tool that can increase the signal-to-noise ratio while maintaining temporal resolution. Combined with LIGHTNING, it adapts to the dynamics of the specimen for the best possible spatial and temporal resolution.

To access functional information, the TauSense technology (1) is a new, straightforward way to access lifetime-based information in confocal imaging. Changes in the fluorescence photon arrival times give an extra layer of information to understand the function of molecules within the cellular environment, to increase image quality, and to expand the number of probes that can be visualized in a specimen. The implementation of different tools (TauConstrast, TauGating, TauScan, and TauSeparation) allows to explore this extra dimension of information at different levels and in a guided way.

In the first part of the webinar we will go through the operating principles behind Photon Counting, LIGHTNING with DSE/AiviaMotion and TauSense using different types of samples. We will show how the different tools are integrated in the STELLARIS confocal platform to get insight into the structure and function from live imaging.

KEYWORDS: Live Imaging, Confocal Imaging, Power Counting, Functional Imaging, CLEM, correlative light and electron microscopy

SPEAKER:

Christophe Leterrier, Aix Marseille Université, CNRS, INP UMR7051, NeuroCyto, Marseille, France

ABSTRACT:

The intricate morphology and molecular identity of axons is maintained for decades, but also continuously adapts to changes in the environment and activity of neurons. Axons fulfill these paradoxical demands thanks to a unique cytoskeletal organization that ensures the coordinated transport, anchoring and mobility of axonal components. In our lab, we use super-resolution microscopy to map the nanoscale architecture actin-based structures within the axon. In the axon initial segment, a key compartment for the maintenance of neuronal polarity, we resolved a highly organized assembly encompassing the periodic actin/spectrin scaffold and its partners: ankyrin, myosin. We have also visualized new actin structures along the axon shaft: rings, hotspots and trails, and are now exploring their molecular organization and functions as well as the role of actin at presynapses. For this, we develop a combination of versatile labeling, correlative live-cell/super-resolution/electron microscopy and quantitative, deep-learning based analysis that allow for high-content, nanoscale interrogation of the axonal architecture.

SPEAKERS:

Flavio Giacobone, Olympus Europa SE & Co. KG
Wei Juan Wong, Olympus Soft Imaging Solutions GmbH

ABSTRACT:

Discover Flexible Slide Scanning

The Olympus Slideview VS200 is a flexible solution for high-throughput and high-quality slide scanning. It has five different observation methods – brightfield, fluorescence, darkfield, phase contrast and polarized light – and can automatically handle a mixture of slide sizes from 26 × 76 mm to 102 × 127 mm. Fluorescence multiplexing is also supported, which makes it possible to create images with up to 32 channels with reliable and precise alignment between channels.

Outstanding Image Quality, Reliable Quantification

For crisp, analysis-ready images, the VS200 captures every detail from slides, across all five observation methods. The optical path has been designed specifically to take advantage of Olympus’ new X Line objectives, which overcome the trade-off between resolution, image flatness and color accuracy – delivering improvements in all three aspects simultaneously. Using true magnifications, from 2X to 100X, it also includes automated oil scanning while the stable and accurate focus of the VS200 gives users confidence when scanning slides for quantification purposes.

A Fluid Workflow for Every User

The VS200 is designed for users of all levels. The basic operation mode enables users to acquire images in as little as three clicks, while the expert mode provides experienced users full control, allowing them to set custom settings to different imaging projects. This is particularly useful if an imaging project requires the acquisitions of different sets of slides over a long timeframe. For ease of use and throughput, the VS200 has a robotic automated slide loader that can handle up to 210 slides, also in mixed sizes. The hot-swap function allows users to change slides during scanning, in case of urgent needs. Efficient working is natural, since the VS200 makes it possible to edit scanned images in parallel with ongoing scans. An automated barcode reader provides reliable capture and recording of information about each slide. The VS200 slide scanner software and hardware can be configured and expanded onsite: going from simple brightfield up to multiplex fluorescence scanning, with deconvolution and AI functions, allows facilities to effortlessly follow the constantly evolving application needs.

KEYWORDS: high-throughput, slide scanning, slide imaging, brain research, drug discovery

SPEAKER:

Ralf Dietzel, Omicron-Laserage Laserprodukte GmbH

ABSTRACT:

Omicron introduces its latest development LaserNest^®, a single-mode desktop laser for microscopy. The presentation also includes information about the multi-wavelength laser light engines LightHUB+ and LightHUB Ultra with up to seven wavelengths.

SPEAKER:

Jan Giesebrecht, Supervisor, Product Applications Specialist, Thermo Fisher Scientific

ABSTRACT:

This live webinar will focus on the visualization and image processing of large 3D image data using the new capabilities of the Amira™ Software solution by Thermo Fisher. The head of the webinar will give you the possibility to learn, through a step-bystep demo, the recently added functions of Amira™ and its new Xplore5D extension. XPlore5D offers the fast and easy exploration of large multi-channel and time-series data. We will also highlight and demonstrate Deep Learning based training and prediction modules.

High-Speed Atomic Force Microscopy and Super-Resolution Optics – Multiparametric Correlative Microscopy Solutions from Bruker Nano Surfaces

SPEAKERS:

Dr. Tanja Neumann, JPK BioAFM, Bruker Nano GmbH,
Dimitar Stamov, JPK BioAFM, Bruker Nano GmbH,

ABSTRACT:

Atomic force microscopy (AFM) currently offers premium spatial resolution of the analysed samples while simultaneously being able to correlate topography and mechanics at near native/physiological imaging conditions. The most recent AFM developments have made it further possible to resolve single molecular dynamic processes on the millisecond scale. In turn, the combination with advanced/customised optics leverages the advantages of immunolabelling techniques for truly correlative microscopy.

During this webinar we will give an overview of correlative microscopy applications featuring fast imaging and advanced super-resolution optical setups. We will also show examples of the accuracy of registering/overlay the AFM and optical images on commercially available DNA origami structures with dimensions below the diffraction limit.

We will further introduce how some of the most recent technological developments have led to unprecedented imaging rates in fluid, setting new milestones in high-speed scanning capabilities. Bruker recently launched the fastest commercially available high-speed AFM (NanoRacer®) enabling real-time visualization of dynamic biological processes taking place on the sub-20-milisecond scale. We will introduce the concept and applications of high-speed imaging for real-time visualization of protein binding kinetics, as well as molecular and cellular dynamics.

Applying Single Molecule Localization Microscopy to Structures Deep Inside the Sample

SPEAKER:

Dr. Clemens Schneider, Sales product Specialist FM EEMEA, Bruker

ABSTRACT:

Single Molecule Localization Microscopy (SMLM) is a very widely used approach for superresolution microscopy. Compared to intensity-based superresolution microscopy images (like in STED or SIM), the localization data obtained in SMLM experiments open new ways for data processing and analysis. Having the precise 3D localization information of individual dye molecules in the sample can be used efficiently for analysis of resolution, spatial distribution, clusters, true colocalization (of clusters) and much more.

In most commercially available systems, all of these advantages are only exploited in regions close to the cover slip, since TIRF (or “dirty-TIRF”) illumination is used for reducing the background coming from out-of-focus planes. The Bruker Vutara VXL system is different in that respect. The dedicated widefield illumination in combination with the biplane detection module opens up the possibility of acquiring SMLM data more than 30 μm inside the sample.

This workshop will explain the technical requirements for performing SMLM deep inside the sample and demonstrates post-acquisition steps for SMLM data processing and analysis.

The Sample Zoo and adequate Light-Sheet Microscope Solutions from Luxendo, Bruker FM

SPEAKER:

Dr. Malte Wachsmuth, Luxendo GmbH, Bruker FM

ABSTRACT:

Light-sheet microscopy (LSFM) is today a well-known and valuable technology to gain new and deep insights of a wide variety of scientific research areas. Sample imaging across scales – with low photo toxicity and low photo bleaching effects at very high speed.

Which LSFM geometry is the most appropriate for your actual sample size, the structure you want to image, and your desired resolution? What are prerequisites to take into account to acquire the most fascinating, high content images?

Which mounting procedures are the most useful for what type of sample? Live samples, fixed samples or cleared samples, they all have specific needs and treats, and these will be addressed.

In this webinar we present an overview of our product portfolio and the key facts regarding lenses, geometries, sample size, mounting techniques, and resolution that characterizes the different specialisations of our light-sheet microscopes.

SPEAKERS:

Raino Ceccarelli, PhD., Head Of Product Development, CrestOptics
Luca Clario, Business Developer Specialist, CrestOptics

ABSTRACT:

The number of biology-driven publications that use Super-Resolution Structured Illumination Microscopy (SR-SIM) has increased significantly in recent years, indicating the usefulness of the technique as a tool for discovery. The increased use of SR-SIM also highlights the limitations of the current approaches. Revealing more biological complexity requires fast simultaneous acquisition of as many labels as possible, deep inside complex specimens. A major handicap of all SR methods is their susceptibility to aberrations, especially when imaging deeper than 10μm, which impacts contrast and resolution. To enable all researchers to overcome this multidimensional challenge, CrestOptics has developed DeepSIM, the first SR-SIM module that is compatible with all standard up-right or inverted microscopes. Despite being a high-end super resolution system (100nm in lateral X,Y and 300nm axial Z resolution), DeepSIM is as easy to operate as a confocal system – it doesn’t require any special probes and researchers can maintain their standard sample preparation protocols which guarantees a seamless evolution from Confocal to SR. We have also equipped the system with robust system calibration, pre-optimized acquisition modalities and automated data processing to make the system easier to use. We are aware that establishing SR-SIM as a ubiquitous tool for routine life science research applications requires much more than just ergonomic design and intuitive handling. With this in mind, we have developed the DeepSIM to be a truly enabling technology. Performance is combined with the flexibility of a modular, expandable system available either as standalone equipment or SR add-on module for CrestOptics X-light V3 Spinning Disk system. To further expand SR-SIM application for the study of structural dynamics and molecular interactions, CrestOptics has dedicated itself to improving live-cell imaging capabilities, despite non-trivial challenges like increasing temporal resolution and adapting to 3D imaging deep inside complex specimens. The webinar will showcase the new DeepSIM SR module and demonstrate how it can help researchers to obtain truly meaningful and contextualized deep data, even from live samples over 50μm thick.

SPEAKERS:

Marit Smeets, Delmic B.V.
Katherine Lau, Delmic B.V.
Guido Ridolfi, Delmic B.V.

ABSTRACT:

Unlocking the power of cryo-ET

Cryo-electron tomography (cryo-ET) is an extremely powerful technique that allows studies of the cellular content at the nano-scale in a near-native state. However, the current cryo-ET workflow is error-prone resulting in damaged, contaminated and devitrified samples that do not result in useful tomography data. In the workflow, there are two main challenges: keeping the sample ice contamination-free and capturing the region of interest (ROI) in the final sample.

To overcome these problems, we developed two workflow solutions called METEOR and CERES to streamline the entire cryo-ET preparation workflow. METEOR is an integrated fluorescent light microscope (FLM) that greatly enhances the ROI targeting inside the FIB/SEM while reducing the number of handling steps required. The CERES Ice Defence System consists of multiple innovative tools that are tailored to minimize ice contamination at different steps of the workflow. In this webinar we will show how using both CERES and METEOR will increase the efficiency of the entire workflow and dramatically increase the amount of high-quality cryo-ET data that can be turned into new biological insights.

Bridging the resolution gap with large-scale EM

Correlative Light Electron Microscopy is a symbiosis of techniques: fluorescence microscopy has a broad range of capabilities such as detecting specific proteins or molecules and monitoring dynamic processes in live material. EM provides data on the structural basis of these processes, by showing all membranes and structures in the sample. Combined, these two are an extremely useful research tool.

The EM part of the workflow is a bottleneck that typically forces investigators to only focus on a much smaller region of interest, compared to what can be observed with Fluorescence Microscopes. This means that unless a substantial amount of time is spent on EM, the technique is primarily qualitative.

Here we demonstrate how FAST-EM, a 64-beam electron microscope designed to drastically increase imaging throughput while maintaining high image quality, is used to collect images at nm resolution from large tissue samples. The improved throughput and automation of FAST-EM enables a shift towards using EM as a fast quantitative analysis tool. Higher throughput drastically shortens the time needed to acquire project data, and opens the door for projects of unprecedented scale, so to closing the resolution gap between EM and FM.

With FAST-EM we can easily capture with EM mode a FOV that is as large as the FM FoV (100 times faster) and collect detailed information on every labeled spot.

SPEAKER:

Bruno Sanguinetti, CTO at Dotphoton

ABSTRACT:

The combination of fast imaging techniques and machine learning are leading to scientific breakthroughs as well as high-impact applications, such as personalized medicine or autonomous driving. They are also generating vast quantities of image data, which is difficult to move around as well as expensive in time, energy and money. Image compression can help, but the usual techniques are not suitable for quantitative data.

Here we present Jetraw, a raw image compressor that preserves the metrological accuracy of raw data, and achieves compression of 8:1 at the speed of 10Gbps networks. This performance is achived through a metrological traceability to the specific image sensor and settings, which gives Jetraw images additional powers, such as physically-accurate data-augmentation and normalization, features that allow to increase, and to test, the reliability of AI algorithms.

SPEAKERS:

Henning Ortkrass*, Marcel Müller*, Anders Kokkvoll Engdahl*, Thomas Huser*,
Gerhard Holst**

*Biomolecular Photonics, Department of Physics – D3, University of Bielefeld, Germany;
**Research Department, PCO AG, Kelheim, Germany

ABSTRACT:

There is a trend in microscopes towards larger field-if-view and larger image circles to be able to receive more information with the recording of one image. For that reason camera manufacturers followed in designing and creating new scientific cameras with larger image sensors and diagonals as well. But, not only the diagonal but even more the pixel size has to be selected properly according to the applied microscope magnification. The relationship will be discussed within this webinar.

Further, it is known that all microscope systems have an optical transfer function or modulation transfer function, and so do cameras. But what is the impact on modern microscope methods like single molecule localization microscopy and structured illumination microscopy? To answer this question, a series of experiments has been done with 4 scientific cameras, which had image sensors with the same pixel size but different MTF. First results are presented within this webinar.

SPEAKER:

Dr. Kirstin Elgass, Business Sector Life Sciences,  ZEISS Research Microscopy Solutions

ABSTRACT:

Since the routine use of fluorescent proteins began in the 1990s, the drive to image live samples has revolutionised our understanding of modern biology. For all live experiments, the most vital consideration is keeping light exposure to an absolute minimum since damage caused by excessive light dose can compromise the integrity of the resulting datasets.

Very often minimising light dosage means reducing the number of acquired images to decrease the length of the timelapse or the frequency of image capture. However, either of these approaches risks missing key events of interest.

Lattice light sheet technology enables 3D acquisition at subcellular resolution and with unprecedented gentleness. First published in 2014, this approach has proven itself to be extremely powerful for long-term studies and homebuilt instruments have generated numerous high impact publications. However, the scope and usability of this approach 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 optimisations for generation of the exciting results.

This has all changed with the release of the ZEISS Lattice Lightsheet 7. This platform provides lattice light sheet imaging but in a convenient, inverted configuration that is compatible with the same samples and sample preparation you would use when imaging with your confocal or spinning disc microscope.

The automatic alignments and easy workflows provided by the inverted Lattice Lightsheet 7 mean that even inexperienced users can now access this cutting-edge approach and capture 3D data of their classically mounted samples 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.