Wiley & ZEISS are proud to present: New Discoveries from the Ultrastructure of Life - Learn How Volume EM Advances Life Science Research

Register for the free webinar series & learn from experts

Intricate ultrastructural information can be delivered by SEM technologies and methods, known collectively as volume EM (vEM). Scientific advancements along with partnerships between the research and commercial communities have seen these methods become both easier to use and more accessible, even to those with little to no experience with electron microscopy.

In this webinar series, we will explore the technological underpinnings of vEM imaging and highlight its growing number of application areas.

We will focus on vEM-specific sample preparation, and then look in detail at individual vEM technologies: Array Tomography Serial Block-Face SEM, and FIB-SEM.

The webinar series will be rounded out with insights into advanced image processing, data analysis, and result visualization capabilities of workflow-oriented software solutions.

Tune in to this series of six webinars and learn how vEM drives new discoveries in neurobiology, cancer research, developmental biology, plant science, and more.

Here are the topics covered in the webinar series:

Webinar #1: An Introduction to volume EM

Webinar #2: Sample Preparation for volume EM

Webinar #3: Easy access to volume EM with Array Tomography

Webinar #4: Automated volume EM with serial block-face imaging

Webinar #5: High-resolution ultrastructure in accurate proportions with FIB-SEM

Webinar #6: Image Processing and Visualization of volume EM data

Note: Webinars 1-6 will be released in Jan-June, 2024 respectively, with 1 releasing per month. Webinar 1&2 are available now, webinar 3 onwards coming soon.

Webinar 1: An Introduction to Volume EM

Speaker:

Kirk Czymmek, Principal Investigator, Director, Advanced Bioimaging Laboratory, Donald Danforth Plant Science Center

Life is inherently three-dimensional (3-D). Thus, approaches that can acquire, visualize, and measure the spatial relationship of organelles, cells, tissues, and organisms allow for greatly improved interpretations and deeper insights for many important biological questions.

Until more recently, capturing cell/tissue volumes at the nanoscale level was a daunting, inefficient, and highly manual process. The development of volume electron microscopy (vEM), a broad collection of imaging strategies that capture hundreds or thousands of consecutive sections in an automated fashion, has extended the power and scope of 3-D cell ultrastructure far beyond traditional transmission electron microscopy (TEM) of individual ultrathin sections. One approach, array tomography (AT), collects a ribbon or “array” of serial ultrathin sections via an ultramicrotome onto a substrate (e.g., grid, glass slide/coverslip or silicon wafer). These sections can be probed with antibodies, stained with heavy metals for contrast, imaged by light, transmission or scanning electron microscopy and archived for future interrogation.

Alternatively, serial block-face scanning electron microscopy (SBF-SEM) employs a diamond knife with an ultramicrotome placed inside a scanning electron microscope which repeatedly removes a thin surface layer from a resin embedded specimen followed by imaging of its rigid block-face. In a related block-face approach, focused ion beam scanning electron microscopy (FIB-SEM) uses an ion beam to remove even thinner layers, enabling the acquisition of high-resolution isotropic 3-D volumes.

This presentation will i) provide an overview of key vEM workflows, how they work and when to use, ii) describe the fundamentals of sample preparation, iii) include examples of biological questions that can be answered, and iv) share community vEM resources to support vEM understanding and success.

Webinar 2: Sample Preparation for Volume EM

Speakers:

Christel Genoud, Ph.D, Senior Lecturer, Faculty of Biology and Medicine & CEO of the Dubochet Center for Imaging. EPFL and Universities of Lausanne, Geneva, and Bern, Switzerland
Corrado Cali, Ph.D, Associate Professor, Department of Neuroscience, University of Turin, Italy
Jean Daraspe, Expert Scientist, Deputy Head, Universite de Lausanne

In this webinar, we aim to explore the impact of sample preparation on various aspects of volumeEM , from sample preparation, to data acquisition and processing. The complexity and breadth of the field preclude a one-size-fits-all approach, yet we aim to shed light on common challenges and necessary considerations. The session is structured into three segments:

First, Christel Genoud will delve into how factors such as fixation, contrast, and embedding type affect the resolution, acquisition speed, and overall quality of the resultant images. Next, Jean Daraspe will present practical examples illustrating the variability in sample mounting and trimming, contingent upon the specific goals of image acquisition and the desired outcomes. He will suggest strategies for targeting regions of interest on a stub or block, and discuss how mounting techniques can influence both charging and imaging conditions. Finally, Corrado Cali will focus on micrograph processing, demonstrating how sample preparation not only affects the interpretation of images but also the ease of reconstruction. He will specifically focus on how fine-tuning image parameters during pre-processing can aid in identifying different cell types, and improves image segmentation tasks.

Overall, this webinar aims to equip participants with tangible examples and insights to critically evaluate and enhance their sample preparation techniques, tailored to their unique research needs.

Webinar 3: Array Tomography: a flexible and accessible volume EM technique

Speakers:

Luke Noon, Ph.D, Principal Investigator, CIPF research institute
Corrado Cali, Ph.D, Associate Professor, Department of Neuroscience, University of Turin, Italy
Jemima Burden, Ph.D, Head of Electron Microscopy, University College London

Array tomography is a volume EM technique where serial ultra-thin sections are cut and collected onto a substrate, and then imaged in an SEM to build up a 3D representation of that sample. Due to the non-destructive nature of the technique, and the stability of the sections on substrate, arrays are amenable to repeated sessions of automated image acquisition, providing unlimited new regions of interest according to the user’s needs and data handling capabilities.

Firstly, Jemima Burden will introduce the Array Tomography workflow, demonstrate how these steps contribute to making it one of the more flexible volume EM techniques, and share how this offers real practical benefits to biomedical and clinical researchers, core facilities and community groups.

Ian White will explain how, with standard equipment that many EM labs will already have, array tomography is a volume EM technique that is genuinely accessible to all on some level and showcase some of the tips and tricks already shared in the growing array tomography community that could help you start accessing the benefits of array tomography for your research now!

Finally, Luke Noon will provide a researcher’s perspective and share how access to this remarkable technique has transformed his lab’s ability to sample liver tissue for the tracing and reconstruction of hepatic nerve endings.

Webinar 4: Automated volume EM with serial block-face imaging

Speaker

Laura Matino, Research fellow, School of Medicine and Surgery, Università degli studi di Milano Bicocca

Complete understanding of biological structures and functions needs three-dimensional imaging to better depict their original view. Serial Block Face Scanning Electron Microscopy (SBF-SEM) offers new opportunities to capture volume information in a slice-and-view automated fashion.

In this webinar, latest breakthroughs in SBF-SEM instrumentation will be shown, focusing on Volutome technology and its improvements in resolution, speed and automation. Next, three different en bloc staining procedures will be considered and compared to speed up tissue preparation while ensuring sample preservation and good resulting contrast. Strategies to maximize the final imaging outcome (i.e., mounting techniques, sample discharge with the focal charge compensation) will be suggested, as well as approaches for 3D reconstruction examined.

& more coming soon…

Webinar speakers

Kirk Czymmek

Principal Investigator, Director, Advanced Bioimaging Laboratory Donald Danforth Plant Science Center

Dr. Kirk Czymmek received his Ph.D. in Botany and Plant Pathology and has over 30 years’ experience dedicated to advanced microscopy techniques including most forms of light, x-ray, electron microscopy and correlative microscopy. His work has focused on developing and applying cutting-edge microscopy tools for imaging cells, tissues, and biomaterials.

Christel Genoud, Ph.D

Senior Lecturer, Faculty of Biology and Medicine & CEO of the Dubochet Center for Imaging EPFL and Universities of Lausanne, Geneva, and Bern, Switzerland

Christel Genoud is an electron microscopist with a background in neuroscience. She studied at the University of Lausanne and got her PhD studying the plasticity of the mouse cortex with electron microscopy under the supervision of Graham Knott. She then went in industry and worked for a company to develop and launch the volumeSEM technique inspired by the microtome developed by W. Denk. In 2008, she joined the Friedrich Miescher Institute in Basel (a Novartis Research Foundation affiliated to the University of Basel) and built an EM facility focuses on volume SEM techniques. From 2016-2020, she managed as well the cryo-EM facility of the FMI shared with Novartis contributing to its creation. Since 2020, she is leading the Em facility of the University of Lausanne. In 2022, she has been nominated CEO of the Dubochet Center for Imaging, a multi-institutional cryo-EM center with platforms in three universities , Lausanne, Geneva and Bern as well as at EPFL.

Corrado Cali, Ph.D.

Associate Professor, Department of Neuroscience University of Turin, Italy

Corrado Calì was trained an electronic engineer at Politecnico di Torino, Italy. He completed his MSc in 2006, in the lab of Henry Markram at EPFL (Lausanne, Switzerland), where he became interested in neuroscience. In the same year he joined the lab of Paola Bezzi at UNIL (University of Lausanne, Lausanne, Switzerland), where he got his doctoral degree in 2012. His research was focused in elucidating the physiological role of astroglial cells in the modulation of synaptic transmission, by releasing neuroactive compounds (so called "gliotranmsission"). After one year as postdoctoral fellow in Graham Knott’s lab, where he enriched and deepend his skills in the state-of-the-art electron microscopy techniques, he joined Pierre Magistretti’s lab in KAUST (King Abdullah University of Science and Technology, Thuwal, Saudi Arabia). Here, he investigated the mechanisms of metabolic support of astrocytes to neurons using morphological and 3D imaging approaches. In particular, he pioneered the use of VR (virtual reality) in neuroscience and is actively exploring scientific visualization approaches for explorative analysis of sparse and dense reconstructions of the CNS from serial-section Electron Microscopy. His interests in 3D visualization and augmented reality led him to establish a company, Intravides (Torino, Italy) with a more clinical focus, developing an Augmented Reality medical device to assist and improve neurosurgical training and practice. Since February 2020, he joined the department of Neuroscience of the University of Torino (Italy), and is now associate professor of Human Anatomy.

Jean Daraspe

Expert Scientist, Deputy Head Universite de Lausanne

Jean obtained his master’s degree in plant sciences at the University of Paris XI in 2005 using confocal microscopy. He acquired his knowledge of electron microscopy after joining the team of Dr. Jean-Marc Verbavatz at the CEA in Saclay, France. Jean joined the EMF in 2008 as a research scientist. He is in charge of all aspects of sample preparation available at the EMF for transmission electron microscopy (TEM) preparation and imaging, using his years of practical experience with biological samples. He is also involved in the development of informatics tools for the automatic image acquisition and analysis of bio-EM samples. Jean trains users in electron microscopy during one-on-one hands-on training sessions and in workshops given by the facility.

Jemima Burden, Ph.D

Head of Electron Microscopy, University College London

Jemima is Head of Electron Microscopy at the Laboratory for Molecular Cell Biology (LMCB) at University College London (UCL). She received her BSc from University of Bath and her PhD from Imperial College London, where her fascination for microscopy began, before specialising in electron microscopy at the LMCB.

Ian White, Ph.D

Deputy Head of Electron Microscopy, LMCB

Ian is Deputy Head of Electron Microscopy at the LMCB. Having completed his PhD at the University of Leeds and a postdoctoral position at the University of Sheffield, Ian moved to UCL where he began to learn the techniques involved in biological electron microscopy, before joining the LMCB as a specialist.

Laura Matino

Research fellow, School of Medicine and Surgery, Università degli studi di Milano Bicocca

Laura Matino received her education in biomedical engineering at Università degli studi di Napoli Federico II where she completed her MSc in 2017. She then joined the Tissue-electronics research group at the Center of Advanced Biomaterials for Healthcare (Istituto Italiano di Tecnologia in Naples) as PhD student, shedding light on novel aspects of neural behavior from the interaction with micro and nanostructured platforms. During her doctoral journey, Laura acquired knowledge of both optical and electron microscopy techniques. Subsequently, she deepened her skills in volume electron microscopy after joining M.D. Guido Cavaletti’s research team at Bicocca School of Medicine and Surgery (Università degli studi di Milano Bicocca), where she optimized biological sample preparation, acquisition and 3D reconstruction of peripheral nervous tissues from serial-section electron microscope imaging.

Luke Noon, Ph.D.

Principal Investigator, CIPF research institute

Luke Noon is principal investigator and head of the Metabolic Growth Signals and Regenerative Medicine Laboratory at the CIPF research institute (Valencia, Spain). His research focuses on how metabolic disease impacts tissue homeostasis and wound healing, with particular emphasis on how changes in peripheral innervation impair regeneration. After training as a zoologist, Luke transitioned to endocrinology and cell/tissue biology where he has developed expertise in Schwann cells, animal models of diabetes and liver disease. He has held postdoctoral positions in the UK (University College London), Spain (CIBERDEM, Valencia) and the USA (Icahn School of Medicine at Mount Sinai, New York). In this webinar, Luke describes how adopting AT-SEM has been a key step in developing a new methodology for mapping the connectivity of peripheral nerves – work performed in close collaboration with Jemima Burden and Prof. Alison Lloyd at the MRC Laboratory for Molecular Cell Biology (UCL, London).

Chris Parmenter, Ph.D.

Editor-in-chief Microscopy and Analysis, Wiley

Chris is a highly-educated chemist with a degree from the University of Hull and an industrial placement at Röhm GmbH in Germany. During his Ph.D. at Warwick University, he synthesized and characterized hydrophobic-hydrophilic block copolymers, and later took a Post-Doc role in electron microscopy. Currently, Chris is a Research Officer at the Nottingham Nanotechnology and Nanoscience Centre, where he is in charge of an FEI Quanta 3D FIB-SEM machine. His research focuses on the self-assembly of nanoscale molecules, particles, devices, and structures.

Register for the free VolumeEM webinar series now

Webinar #1: An Introduction to Volume EM

Webinar #2: Sample Preparation for Volume