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Phage display-derived alpaca nanobodies as potential therapeutics for Naja atra snake envenomation

Applied and Environmental Microbiology 2024年第8期90卷 1页

作者： Wang, Wei-Chu Chang, Jungshan Lee, Chi-Hsin Chiang, Yu-Wei Leu, Sy-Jye Mao, Yan-Chiao Chiang, Jen-Ron Yang, Chun-Kai Wu, Chao-Jung Yang, Yi-Yuan Ph.D. Program in Medical Biotechnology College of Medical Science and Technology Taipei Medical University Taipei Taiwan Graduate Institute of Medical Sciences College of Medicine Taipei Medical University Taipei Taiwan International Master/Ph.D. Program in Medicine College of Medicine Taipei Medical University Taipei Taiwan International Ph.D. Program for Cell Therapy and Regeneration Medicine College of Medicine Taipei Medical University Taipei Taiwan School of Medical Laboratory Science and Biotechnology College of Medical Science and Technology Taipei Medical University Taipei Taiwan Core Laboratory of Antibody Generation and Research Taipei Medical University Taipei Taiwan Department of Medical Research Taipei Veterans General Hospital Taipei Taiwan Department of Biology and Anatomy National Defense Medical Center Taipei Taiwan Foundation for Poison Control Taipei Taiwan Department of Microbiology and Immunology School of Medicine College of Medicine Taipei Medical University Taipei Taiwan School of Medicine National Defense Medical Centre Taipei Taiwan Department of Post-Baccalaureate Medicine College of Medicine National Chung Hsing University Taichung Taiwan Division of Clinical Toxicology Department of Emergency Medicine Taichung Veterans General Hospital Taichung Taiwan Bioproduction Plants National Institute of Infectious Diseases and Vaccinology National Health Research Institutes Miaoli Taiwan Interdisciplinary Research on Ecology and Sustainability National Dong Hwa University Hualien Taiwan

Naja atra, the Chinese cobra, is a major cause of snake envenomation in Asia, causing hundreds of thousands of clinical incidents annually. The current treatment, horse serum-derived antivenom, has unpredictable side effects and presents manufacturing challenges. This study focused on developing new-generation snake venom antidotes by using microbial phage display technology to derive nanobodies from an alpaca immunized with attenuated N. atra venom. Following confirmation of the immune response in the alpaca, we amplified VHH genes from isolated peripheral blood mononuclear cells and constructed a phage display VHH library of 1.0 × 107 transformants. After four rounds of biopanning, the enriched phages exhibited increased binding activity to N. atra venom. Four nanobody clones with high binding affinities were selected: aNAH1, aNAH6, aNAH7, and aNAH9. Specificity testing against venom from various snake species, including two Southeast Asian cobra species, revealed nanobodies specific to the genus Naja. An in vivo mouse venom neutralization assay demonstrated that all nanobodies prolonged mouse survival and aNAH6 protected 66.6% of the mice from the lethal dosage. These findings highlight the potential of phage display-derived nanobodies as valuable antidotes for N. atra venom, laying the groundwork for future applications in snakebite treatment. Copyright © 2024 American Society for Microbiology. All Rights Reserved.

关键词： Mammals

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Lotus seed skin proanthocyanidin extract exhibits potent antioxidant property via activation of the Nrf2-ARE pathway

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生物化学与生物物理学报（英文版） 2019年第1期51卷 31-40页

作者： Tao Li Qili Li Weiguo Wu Yong Li De-xing Hou Hua Xu Baodong Zheng Shaoxiao Zeng Yang Shan Xiangyang Lu Fangming Deng Si Qin Core Research Program 1515 Key Laboratory for Food Science and Biotechnology of Hunan Province College of Food Science and Technology Hunan Agricultural University Changsha 410128 China Hunan Agricultural Product Processing Institute Hunan Academy of Agricultural Sciences Changsha 410125 China Core Research Program 1515 Key Laboratory for Food Science and Biotechnology of Hunan Province College of Food Science and Technology Hunan Agricultural University Changsha 410128 China The United Graduate School of Agricultural Sciences Faculty of Agriculture Kagoshima University Kagoshima 890-0065Japan Department of Pediatrics Steele Children's Research Center University of Arizona Tucson 85724 USA Department of Food Safety College of Food Science Fujian Agriculture and Forestry University Fuzhou 350002 China Core Research Program 1515 Key Laboratory for Food Science and Biotechnology of Hunan Province College of Food Science and Technology Hunan Agricultural University Changsha 410128 China The United Graduate School of Agricultural Sciences Faculty of Agriculture Kagoshima University Kagoshima 890-0065Japan Department of Pediatrics Steele Children's Research Center University of Arizona Tucson 85724 USA

Lotus seed is well known as traditional food and medicine,but its skin is usually *** studies have shown that lotus seed skin contains a high concentration of proanthocyanidins that have multi-functions,such as antioxidation,anti-inflammation,and anti-cancer *** the present study,we aimed to isolate and purify the proanthocyanidins from lotus seed skin by acetone extraction and rotary evaporation,identify their chemical structures by HPLC-MS-MS and NMR,and further investigate the antioxidant properties of the extract purified by macroporous resin (PMR) from lotus seed skin both in vitro and in *** results showed that PMR mainly contained oligomeric proanthocyanidins,especially dimeric procyanidin B1 (PB1),procyanidin B2 and procyanidin *** it had limited ability to directly scavenge radicals in vitro,PMR could significantly enhance the expressions of antioxidant proteins via activation of nuclear factor-E2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway in HepG2 *** data revealed that PB1,a major component in PMR,stabilized Nrf2 by inhibiting the ubiquitination of Nrf2,which led to subsequent activation of the Nrf2-ARE pathway,including the enhancements of Nrf2 nuclear translocation,Nrf2-ARE binding and ARE transcriptional ***,the in vivo results in high fat diet-induced mice further verified the powerful antioxidant property of *** results revealed that lotus seed skin is a promising resource for functional food development.

关键词： lotus seed skin extract proanthocyanidin Nrf2-ARE pathway antioxidant activity

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Harnessing microalgae to mitigate the environmental impact of the cement industry: Emission reduction and bio-cement production

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Carbon Capture science and technology 2025年 16卷

作者： Le, Ve Van Esterhuizen, Maranda Tran, Quynh-Giao Yun, Jin-Ho Jung, Man-Young Lee, Sang-Ah Faculty of Biotechnology College of Applied Life Sciences Jeju National University 102 Jejudaehak-Ro Jeju 63243 South Korea Interdisciplinary Graduate Program in Advance Convergence Technology and Science Jeju National University 102 Jejudaehak-ro Jeju 63243 South Korea Bio-Health Materials Core-Facility Center Jeju National University 102 Jejudaehak-ro Jeju 63243 South Korea Department of Biology Education Jeju National University Jeju 63243 South Korea University of Helsinki Ecosystems and Environment Research Programme Faculty of Biological and Environmental Sciences Niemenkatu 73 Lahti 15140 Finland Cell factory Research Centre Korea Research Institute of Bioscience & Biotechnology 125 Gwahak-ro Yuseong-gu Daejeon 34141 South Korea Department of Environmental Biotechnology KRIBB School of Biotechnology University of Science & Technology Daejeon 34113 South Korea Department of Integrative Biotechnology Sungkyunkwan University Suwon 16419 South Korea

The cement industry has been the cornerstone of economic development since the Industrial Revolution. However, the calcination process used to produce cement raw materials releases various pollutants, such as carbon dioxide, nitrogen oxides, and sulfur oxides, which are key contributors to global warming. Therefore, innovative technologies are urgently needed to minimize the environmental impact of the cement industry. In recent years, microalgae have gained attention because of their advantages in recovering resources and reducing the impacts of industrial pollutants. The use of microalgae to address the environmental challenges associated with cement production remains largely overlooked in current reviews. This review summarizes the recent advances in the utilization of microalgae to address the environmental challenges posed by the cement industry. Microalgae offer two main pathways for environmental mitigation: the (i) production of bio-cement as an eco-friendly alternative for construction and (ii) conversion of cement flue gases into biomass feedstock with high-value downstream applications. We also identify existing challenges and propose an integrative “Microalgae–Bacteria Consortium” system as a sustainable strategy for the significant reduction of the environmental footprint of cement production. This approach has the potential to transform the cement industry into a more sustainable and eco-friendlier sector. Overall, microalgae provide an innovative and sustainable platform to revolutionize the cement industry into a greener sector, supporting the global shift toward a low-carbon, circular economy. © 2025 The Author(s)

关键词： Bio-cement Cement calcination Flue gas Greenhouse gas Microalgae Microalgae–bacteria consortium Sustainable construction

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Graphene Acid Quantum Dots: A Highly Active Multifunctional Carbon Nano Material that Intervene in the Trajectory Towards Neurodegeneration

SSRN

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SSRN 2024年

作者： ElMorsy, Sherin M. Gutierrez, Denisse A. Valdez, Salvador Kumar, Jyotish Aguilera, Renato J. Noufal Sanad, Mohamed Sarma, Hemen Chinnam, Sampath Narayan, Mahesh The Environmental Science & Engineering Program The University of Texas at El Paso El PasoTX79968 United States Cellular Characterization and Biorepository Core Facility Border Biomedical Research Center Department of Biological Sciences College of Science The University of Texas at El Paso 500 West University Avenue El PasoTX79968-0519 United States Department of Chemistry and Biochemistry The University of Texas at El Paso 500 West University Avenue El PasoTexas79968 United States Department of Chemical Engineering Hampton University HamptonVA23668 United States Assam Kokrajhar783370 India Department of Chemistry M.S. Ramaiah Institute of Technology Karnataka Bengaluru560054 India Autonoumous Institution Affiliated to Visvesvaraya Technological University Belgaum India

Carbon dots (CDs) are carbon nano materials (CNMs) that find use across several biological applications because of their water solubility, biocompatible nature, eco-friendliness, and ease of synthesis. Additionally, their physiochemical properties can be chemically tuned for further optimization towards specific applications. Here, we investigate the efficacy of C70-derived Graphene Acid Quantum Dots (GAQDs) in mitigating the transformation of soluble, monomeric Hen Egg-White Lysozyme (HEWL) to mature fibrils during its amyloidogenic trajectory. Our findings reveal that GAQDs exhibit dose-dependent inhibition of HEWL fibril formation (up to 70% at 5 mg/mL) without affecting mitochondrial membrane potential or inducing apoptosis at the same density. Furthermore, GAQDs scavenged reactive oxygen species (ROS);achieving a 50% reduction in ROS levels at a mere 100 µg/mL when exposed to a standard free radical generator. GAQDs were not only found to be biocompatible with a human neuroblastoma-derived SHSY-5Y cell line but also rescued the cells from rotenone-induced apoptosis. The GAQD-tolerance of SHSY-5Y cells coupled with their ability to restitute cells from rotenone-dependent apoptosis, when taken in conjunction with the biocompatibility data, indicate that GAQDs possess neuroprotective potential. The data position this class of CNMs as promising candidates for resolving aberrant cellular outputs that associate with the advent and progress of multifactorial neurodegenerative disorders including Parkinson's (PD) and Alzheimer’s diseases (AD) wherein environmental causes are implicated (95% etiology). The data suggest that GAQDs are a multifunctional carbon-based sustainable nano-platform at the intersection of nanotechnology and neuroprotection for advancing green chemistry-derived, sustainable healthcare solutions. © 2024, The Authors. All rights reserved.

关键词： Graphene

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Combinatorial Engineered Immuno-Fragments Targeting SARS-CoV-2 Nucleocapsid Protein for Epitope Complementarity

SSRN

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SSRN 2024年

作者： Dlamini, Nhlanhla Benedict Lee, Chi-Hsin Wang, Wei-Chu Huang, Peng-Nien Leu, Sy-Jye Tseng, Sung-Nien Shih, Shin-Ru Chiang, Liao-Chun Mao, Yan-Chiao Tsai, Bor-Yu Yang, Yi-Yuan Ph.D. Program in Medical Biotechnology College of Medical Science and Technology Taipei Medical University Taipei110301 Taiwan School of Medical Laboratory Science and Biotechnology College of Medical Science and Technology Taipei Medical University Taipei110301 Taiwan Research Center for Emerging Viral Infections Chang Gung University Taoyuan333423 Taiwan Division of Infectious Diseases Department of Pediatrics Linkou Chang Gung Memorial Hospital Taoyuan333423 Taiwan Graduate Institute of Medical Sciences College of Medicine Taipei Medical University Taipei110301 Taiwan Department of Microbiology and Immunology School of Medicine College of Medicine Taipei Medical University Taipei110301 Taiwan Department of Laboratory Medicine Linkou Chang Gung Memorial Hospital Taoyuan333423 Taiwan College of Life Sciences National Tsing Hua University Hsinchu300040 Taiwan Division of Clinical Toxicology Department of Emergency Medicine Taichung Veterans General Hospital Taichung407219 Taiwan Navi Bio-Therapeutics Inc. Taipei10351 Taiwan Core Laboratory of Antibody Generation and Research Taipei Medical University Taipei110301 Taiwan

Recombinant avian phage display immunoglobulin Y derivative single chain variable fragment (IgY-scFv) antibodies have evolved as robust single-chain antigen recognizing IgY analogues due to the advent of immuno-nanotechnology and other immunomolecular biotechnology applications. This academic study reports on the engineering and characterization of severe acute respiratory syndrome coronavirus 2 nucleocapsid (SARS-CoV-2 N) monoaffinity recombinant chicken IgY derivative single chain variable fragment (IgY-scFv) antibody fragments as versatile tools for surrogate immunological identification of the SARS-CoV-2 virus nuclear biomarker. To improve the immunopanning selection for specific binders, layer hens were immunised with the SARS-CoV-2 nucleocapsid protein to pre-enrich higher affinity B cell gene repertoires prior to library construction. Pre-enrichment humoral egg yolk polyclonal IgY antibodies were isolated following the PEG-8000 precipitation protocol. An experimental based modelling approach was applied in order to create IgY derivative N protein single chain variable fragments (IgY-scFv N) M13KO7 phage antibody libraries, an immune phage display library constructed system that supported the generation of functional anti-SARS-CoV-2 N protein IgY-scFv affinity monospecific fragment immunomolecules. After the immunopanning enrichment selection process three distinct positive sequence family IgY-scFv fragments obtained demonstrated epitope specific affinity (micromolar range kinetics) towards the targeted recombinant SARS-CoV-2 N protein and also positively identified native SARS-CoV-2 (COVID-19) N protein in formalin-fixed paraffin-embedded (FFPE) 293T cell pellets. © 2024, The Authors. All rights reserved.

关键词： Coronavirus

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Effective theory for universal seesaw model and FCNC

arXiv

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arXiv 2022年

作者： Morozumi, Takuya Adam, Apriadi Salim Kawamura, Yuta Panuluh, Albertus Hariwangsa Shimizu, Yusuke Yamamoto, Kei Physics Program Graduate School of Advanced Science and Engineering Hiroshima University Higashi-Hiroshima739-8526 Japan Core of Research for the Energetic Universe Hiroshima University Higashi-Hiroshima739-8526 Japan South Tangerang15314 Indonesia Mie Prefecture Yokkaichi Japan Department of Physics Education Sanata Dharma University Paingan Maguwohardjo Sleman Yogyakarta55282 Indonesia Department of Global Environment Studies Hiroshima Institute of Technology Hiroshima731- 5193 Japan

We study the quark sector of the universal seesaw model with SU(2)L × SU(2)R × U(1). The model incorporates the seesaw mechanism with the vector-like quarks (VLQs). The purpose of this work is to study the model with the effective theory. After integrating the heavy five VLQs, we derive the effective theory with four up-type quark and three down type quark. In this work, the FCNC of Z boson for top quark and top′ quark is derived. © 2022, CC BY.

关键词： Bosons

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Galaxy clusters at z ∼ 1 imaged by ALMA with the Sunyaev–Zel’dovich effect

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Publications of the Astronomical Society of Japan 2023年第2期75卷 311-337页

作者： Kitayama, Tetsu Ueda, Shutaro Okabe, Nobuhiro Akahori, Takuya Hilton, Matt Hughes, John P. Ichinohe, Yuto Kohno, Kotaro Komatsu, Eiichiro Lin, Yen-Ting Miyatake, Hironao Oguri, Masamune Sifón, Cristóbal Takakuwa, Shigehisa Takizawa, Motokazu Tsutsumi, Takahiro van Marrewijk, Joshiwa Wollack, Edward J. Department of Physics Toho University 2-2-1 Miyama Chiba Funabashi274-8510 Japan No. 1 Section 4 Roosevelt Road Taipei10617 Taiwan Institute of Astronomy National Tsing Hua University Hsinchu30013 Taiwan Department of Physics Hiroshima University 1-3-1 Kagamiyama Higashi-Hiroshima Hiroshima739-8526 Japan Astrophysical Science Center Hiroshima University 1-3-1 Kagamiyama Higashi-Hiroshima Hiroshima739-8526 Japan Core Research for Energetic Universe Department of Physics Hiroshima University 1-3-1 Kagamiyama Higashi-Hiroshima Hiroshima739-8526 Japan Physics Program Graduate School of Advanced Science and Engineering Hiroshima University 1-3-1 Kagamiyama Higashi-Hiroshima Hiroshima739-8526 Japan Mizusawa VLBI observatory National Astronomical Observatory of Japan 2-21-1 Osawa Mitaka Tokyo181-8588 Japan Operation Division SKA Observatory Jodrell Bank Lower Withington MacclesfieldSK11 9DL United Kingdom Astrophysics Research Centre University of KwaZulu-Natal Westville Campus Durban4041 South Africa School of Mathematics Statistics & Computer Science University of KwaZulu-Natal Westville Campus Durban4041 South Africa Wits Centre for Astrophysics School of Physics University of the Witwatersrand Private Bag 3 Johannesburg2050 South Africa Department of Physics and Astronomy Rutgers The State University of New Jersey PiscatawayNJ08854-8019 United States Department of Physics Rikkyo University 3-34-1 Nishi-Ikebukuro Toshima-ku Tokyo171-8501 Japan Institute of Astronomy School of Science The University of Tokyo 2-21-1 Osawa Mitaka Tokyo181-0015 Japan Research Center for the Early Universe School of Science The University of Tokyo 7-3-1 Hongo Bunkyo Tokyo113-0033 Japan Max-Planck-Institut für Astrophysik Karl-Schwarzschild Str. 1 GarchingD-85741 Germany The University of Tokyo Institutes for Advanced Study The University of Tokyo 5-1-5 Kashiwanoha Chiba Kashiwa277-8583 Japan Nagoya University Furo-cho Chikusa-

We present high angular resolution measurements of the thermal Sunyaev–Zel’dovich effect (SZE) toward two galaxy clusters, RCS J2319+0038 at z = 0.9 and HSC J0947−0119 at z = 1.1, by the Atacama Large Millimeter/submillimeter Array (ALMA) in Band 3. They are supplemented with available Chandra X-ray data, optical data taken by Hyper Suprime-Cam on Subaru, and millimeter-wave SZE data from the Atacama Cosmology Telescope. Taking into account departures from spherical symmetry, we have reconstructed non-parametrically the inner pressure profile of two clusters as well as electron temperature and density profiles for RCS J2319+0038. This is one of the first such measurements for an individual cluster at z 0.9. We find that the inner pressure profile of both clusters is much shallower than that of local cool-core clusters. Our results consistently suggest that RCS J2319+0038 hosts a weak cool core, where radiative cooling is less significant than in local cool cores. On the other hand, HSC J0947−0119 exhibits an even shallower pressure profile than RCS J2319+0038 and is more likely to be a non-cool-core cluster. The SZE centroid position is offset by more than 140 h−170 kpc from the peaks of galaxy distribution in HSC J0947−0119, suggesting a stronger influence of mergers in this cluster. We conclude that these distant clusters are at a very early stage of developing the cool cores typically found in clusters at lower redshifts. © The Author(s) 2023. Published by Oxford University Press on behalf of the Astronomical Society of Japan.

关键词： Millimeter waves

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Corrigendum to “Socio-economic status and occupational mobility of China’s Fishery Population: A quantitative analysis based on social-survey data” [Fish. Res. 285(2025) 107362]

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Fisheries research 2025年 285卷

作者： Yi Huang Ulf Dieckmann Mikko Heino Coastal and Ocean Management Institute Xiamen University Xiamen 361102 China Evolution and Ecology Program & Advancing Systems Analysis Program International Institute for Applied Systems Analysis (IIASA) Schlossplatz 1 Laxenburg A2361 Austria Complexity Science and Evolution Unit Okinawa Institute of Science and Technology Graduate University (OIST) 1919-1 Tancha Onna Kunigami Okinawa 904-0495 Japan Department of Evolutionary Studies of Biosystems The Graduate University for Advanced Studies (Sokendai) Hayama Kanagawa 240-0193 Japan Department of Biological Sciences University of Bergen Box 7803 Bergen N-5020 Norway Institute of Marine Research Box 1870 Nordnes Bergen N-5817 Norway

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Modulus τ linking leptonic CP violation to baryon asymmetry in A4 modular invariant flavor model

arXiv

引用

arXiv 2021年

作者： Okada, Hiroshi Shimizu, Yusuke Tanimoto, Morimitsu Yoshida, Takahiro Asia Pacific Center for Theoretical Physics Pohang37673 Korea Republic of Department of Physics Pohang University of Science and Technology Pohang37673 Korea Republic of Physics Program Graduate School of Advanced Science and Engineering Hiroshima University Higashi-Hiroshima739-8526 Japan Core of Research for the Energetic Universe Hiroshima University Higashi-Hiroshima739-8526 Japan Department of Physics Niigata University Niigata950-2181 Japan

We propose an A4 modular invariant flavor model of leptons, in which both CP and modular symmetries are broken spontaneously by the vacuum expectation value of the modulus τ. The value of the modulus τ is restricted by the observed lepton mixing angles and lepton masses for the normal hierarchy of neutrino masses. The predictive Dirac CP phase δCP is in the ranges [0◦, 50◦], [170◦, 175◦] and [280◦, 360◦] for Re [τ] ◦, 80◦], [185◦, 190◦] and [310◦, 360◦] for Re [τ] > 0. The sum of three neutrino masses is predicted in [60, 84] meV, and the effective mass for the 0νββ decay is in [0.003, 3] meV. The modulus τ links the Dirac CP phase to the cosmological baryon asymmetry (BAU) via the leptogenesis. Due to the strong wash-out effect, the predictive baryon asymmetry YB can be at most the same order of the observed value. Then, the lightest right-handed neutrino mass is restricted in the range of M1 = [1.5, 6.5]×1013 GeV. We find the correlation between the predictive YB and the Dirac CP phase δCP. Only two predictive δCP ranges, [5◦, 40◦] (Re [τ] > 0) and [320◦, 355◦] (Re [τ] Copyright © 2021, The Authors. All rights reserved.

关键词： Neutrons

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Volume electron microscopy (vol 2, 51, 2022)

NATURE REVIEWS METHODS PRIMERS

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NATURE REVIEWS METHODS PRIMERS 2022年第1期2卷 1页

作者： Peddie, Christopher J. Genoud, Christel Kreshuk, Anna Meechan, Kimberly Micheva, Kristina D. Narayan, Kedar Pape, Constantin Parton, Robert G. Schieber, Nicole L. Schwab, Yannick Titze, Benjamin Verkade, Paul Weigel, Aubrey Collinson, Lucy M. Electron Microscopy Science Technology Platform The Francis Crick Institute London UK Electron Microscopy Facility Faculty of Biology and Medicine University of Lausanne Lausanne Switzerland Cell Biology and Biophysics Unit European Molecular Biology Laboratory Heidelberg Germany Department of Molecular and Cellular Physiology Stanford University Palo Alto CA USA Center for Molecular Microscopy Center for Cancer Research National Cancer Institute National Institutes of Health Bethesda MD USA Cancer Research Technology Program Frederick National Laboratory for Cancer Research Frederick MD USA The Institute for Molecular Bioscience The University of Queensland Brisbane Queensland Australia Centre for Microscopy and Microanalysis The University of Queensland Brisbane Queensland Australia Cell Biology and Biophysics Unit/ Electron Microscopy Core Facility European Molecular Biology Laboratory Heidelberg Germany Nanosurf AG Liestal Switzerland School of Biochemistry University of Bristol Bristol UK Janelia Research Campus Howard Hughes Medical Institute Ashburn VA USA

Life exists in three dimensions, but until the turn of the century most electron microscopy methods provided only 2D image data. Recently, electron microscopy techniques capable of delving deep into the structure of cells and tissues have emerged, collectively called volume electron microscopy (vEM). Developments in vEM have been dubbed a quiet revolution as the field evolved from established transmission and scanning electron microscopy techniques, so early publications largely focused on the bioscience applications rather than the underlying technological breakthroughs. However, with an explosion in the uptake of vEM across the biosciences and fast-paced advances in volume, resolution, throughput and ease of use, it is timely to introduce the field to new audiences. In this Primer, we introduce the different vEM imaging modalities, the specialized sample processing and image analysis pipelines that accompany each modality and the types of information revealed in the data. We showcase key applications in the biosciences where vEM has helped make breakthrough discoveries and consider limitations and future directions. We aim to show new users how vEM can support discovery science in their own research fields and inspire broader uptake of the technology, finally allowing its full adoption into mainstream biological imaging.

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