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University of Birmingham

36 ARTICLES PUBLISHED IN JoVE

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Biology

In situ Subcellular Fractionation of Adherent and Non-adherent Mammalian Cells
Anyaporn Sawasdichai 1, Hsin-Tien Chen 1, Nazefah Abdul Hamid 1, Padma-Sheela Jayaraman 2, Kevin Gaston 1
1Department of Biochemistry, School of Medical Sciences, University of Bristol, 2Division of Immunity and Infection, School of Medicine, University of Birmingham

In situ subcellular fractionation of mammalian cells on microscope coverslips allows the visualisation of protein localisation.

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Biology

Techniques for Imaging Ca2+ Signaling in Human Sperm
Katherine Nash 1, Linda Lefievre 2, Ruben Peralta-Arias 1, Jennifer Morris 1, Aduen Morales-Garcia 1, Tom Connolly 2, Sarah Costello 1, Jackson C. Kirkman-Brown 3, Stephen J. Publicover 1
1School of Biosciences, University of Birmingham, 2School of Medicine, University of Birmingham, 3Centre for Human Reproductive Science, Birmingham Women’s Hospital

Stimulus-evoked [Ca2+]i signals of individual human sperm are assessed. Motile cells are loaded with Ca2+-sensitive fluorescent dye (AM-ester method) and immobilised in a perfusable chamber. Cells are imaged by time-lapse fluorescence microscopy and stimulated via the perfusing medium. Responses of single cells (or regions) are analysed offline using Excel.

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Biology

Adult and Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells
Deborah Merrick 1, Hung-Chih Chen 1, Dean Larner 1, Janet Smith 1
1School of Biosciences, University of Birmingham

The micro-dissected explants technique is a robust and reliable method for isolating proliferative skeletal muscle cells from juvenile, adult or embryonic muscles as a source of skeletal muscle stem cells. Uniquely, these cells have been clonally derived to produce skeletal muscle stem cell lines used for in vivo transplantation.

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JoVE Journal

The Generation of Higher-order Laguerre-Gauss Optical Beams for High-precision Interferometry
Ludovico Carbone 1, Paul Fulda 1, Charlotte Bond 1, Frank Brueckner 1, Daniel Brown 1, Mengyao Wang 1, Deepali Lodhia 1, Rebecca Palmer 1, Andreas Freise 1
1School of Physics and Astronomy, University of Birmingham

Large laser-interferometers are being constructed to create a new type of astronomy based on gravitational waves. Their sensitivities, as for many other high-precision experiments, are approaching fundamental noise limits such as the atomic vibration of their components. We are pioneering technologies to overcome these limits using novel laser beam shapes.

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Immunology and Infection

Generation of Lymph Node-fat Pad Chimeras for the Study of Lymph Node Stromal Cell Origin
Cecile Benezech 1,2, Jorge H. Caamano 1
1School of Immunity and Infection, IBR-MRC Centre for Immune Regulation, College of Medical and Dental Sciences, University of Birmingham, 2Centre for Cardiovascular Sciences, University of Edinburgh

Generation of lymph node/fat pad chimeras for the study of lymph node stromal cell origin is described. The method involves the isolation of lymph nodes from newborn mice and embryonic fat pads, the generation of chimeric lymph node-fat pads, and their transfer under the kidney capsule of a host mouse.

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Immunology and Infection

A Flow Adhesion Assay to Study Leucocyte Recruitment to Human Hepatic Sinusoidal Endothelium Under Conditions of Shear Stress
Shishir Shetty 1, Christopher J. Weston 1, David H. Adams 1, Patricia F. Lalor 1
1NIHR Biomedical Research Unit, Centre for Liver Research, School of Immunity and Infection, University of Birmingham

Leucocyte recruitment to the liver occurs within the specialized channels of the hepatic sinusoids which are lined by unique hepatic sinusoidal endothelial cells. Phase contrast microscopy of leucocyte recruitment across human hepatic sinusoidal endothelium under conditions of physiological shear stress can facilitate the elucidation of the molecular mechanisms which underlie this process.

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Biology

Reduced-gravity Environment Hardware Demonstrations of a Prototype Miniaturized Flow Cytometer and Companion Microfluidic Mixing Technology
William S. Phipps *1, Zhizhong Yin *1, Candice Bae 1, Julia Z. Sharpe 1, Andrew M. Bishara 2, Emily S. Nelson 3, Aaron S. Weaver 3, Daniel Brown 4, Terri L. McKay 3, DeVon Griffin 3, Eugene Y. Chan 1
1DNA Medicine Institute, 2Harvard Medical School, 3NASA Glenn Research Center, 4ZIN Technologies

Spaceflight blood diagnostics need innovation. Few demonstrations have been published illustrating in-flight, reduced-gravity health diagnostic technology. Here we present a method for construction and operation of a parabolic flight test rig for a prototype point-of-care flow-cytometry design, with components and preparation strategies adaptable to other setups.

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Behavior

Modulating Cognition Using Transcranial Direct Current Stimulation of the Cerebellum
Paul A. Pope 1
1School of Psychology, University of Birmingham

Transcranial direct current stimulation (tDCS) over the cerebellum exerts a remote effect on the prefrontal cortex, which can modulate cognition and performance. This was demonstrated using two information-processing tasks of varying complexity, whereby only cathodal tDCS improved performance when the task was difficult, but not easy.

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Immunology and Infection

Analyzing the Effects of Stromal Cells on the Recruitment of Leukocytes from Flow
Hafsa Munir 1,2, G. Ed Rainger 1,2, Gerard B. Nash 1,2, Helen McGettrick 2,3
1School of Clinical and Experimental Medicine, University of Birmingham, 2College of Medical and Dental Sciences, University of Birmingham, 3School of Immunity and Infection, University of Birmingham

The ability of inflamed endothelium to recruit leukocytes from flow is regulated by mesenchymal stromal cells. We describe two in vitro models incorporating primary human cells that can be used to assess neutrophil recruitment from flow and examine the role that mesenchymal stromal cells play in regulating this process.

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Immunology and Infection

Biomimetic Materials to Characterize Bacteria-host Interactions
Daniel H. Stones *1, Fitua Al-Saedi *1, Diana Vaz 1, Nicolas Perez-Soto 1, Anne M. Krachler 1
1Institute of Microbiology and Infection, School of Biosciences, University of Birmingham

Bacterial attachment to host cells is a key step during host colonization and infection. This protocol describes the generation of polymer-coupled recombinant adhesins as biomimetic materials which allow analysis of the contribution of individual adhesins to these processes, independent of other bacterial factors.

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Environment

Dispersion of Nanomaterials in Aqueous Media: Towards Protocol Optimization
Inder Kaur 1, Laura-Jayne Ellis 1, Isabella Romer 1, Ratna Tantra 2, Marie Carriere 3,4, Soline Allard 5, Martine Mayne-L'Hermite 5, Caterina Minelli 6, Wolfgang Unger 7, Annegret Potthoff 8, Steffi Rades 7, Eugenia Valsami-Jones 1
1School of Geography, Earth and Environmental Sciences, University of Birmingham, 2Analytical Science, National Physical Laboratory, 3INAC-LCIB, Université Grenoble Alpes, 4CEA, INAC-SyMMES, 5NIMBE, CEA, CNRS, Université Paris-Saclay, CEA Saclay, 6Chemical, Medical and Environmental Science, National Physical Laboratory, 7BAM Division 6.1 'Surface Analysis and Interfacial Chemistry', BAM Federal Institute for Materials Research and Testing, 8Fraunhofer Institute for Ceramic Technologies and Systems

Here, we present a step-wise protocol for the dispersion of nanomaterials in aqueous media with real-time characterization to identify the optimal sonication conditions, intensity, and duration for improved stability and uniformity of nanoparticle dispersions without impacting the sample integrity.

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Environment

Resurrection of Dormant Daphnia magna: Protocol and Applications
Maria Cuenca Cambronero 1, Luisa Orsini 1
1Environmental Genomics Group, School of Biosciences, University of Birmingham

Long-term studies are essential to understanding the process of evolution and the mechanisms of adaptation. Generally, these studies require commitments beyond the life-time of researchers. Here, a powerful method is described that dramatically advances state-of-the-art data collection to generate longitudinal data in natural systems.

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Immunology and Infection

Stenosis of the Inferior Vena Cava: A Murine Model of Deep Vein Thrombosis
Holly Payne 1, Alexander Brill 1
1Institute of Cardiovascular Sciences, College of Medical and Dental Sciences, University of Birmingham

We describe here stenosis in the inferior vena cava as a murine model of deep vein thrombosis. This model recapitulates blood flow restriction, one of the major triggers of venous thrombosis in humans.

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Medicine

High-Throughput Analysis of Optical Mapping Data Using ElectroMap
Christopher O’Shea 1,2,3, Andrew P. Holmes 1,4, Ting Y. Yu 1, James Winter 1, Simon P. Wells 1, Beth A. Parker 1, Dannie Fobian 1, Daniel M. Johnson 1, Joao Correia 5, Paulus Kirchhof 1, Larissa Fabritz 1, Kashif Rajpoot 3, Davor Pavlovic 1
1Institute of Cardiovascular Sciences, University of Birmingham, 2EPSRC Centre for Doctoral Training in Physical Sciences for Health, School of Chemistry, University of Birmingham, 3School of Computer Science, University of Birmingham, 4Institute of Clinical Sciences, University of Birmingham, 5Institute of Microbiology and Infection, School of Biosciences, University of Birmingham

This protocol describes the setup and use of ElectroMap, a MATLAB-based open-source software platform for analysis of cardiac optical mapping data. ElectroMap provides a versatile high-throughput tool for analysis of optical mapping voltage and calcium datasets across a wide range of cardiac experimental models.

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Developmental Biology

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
Kinga Winczura 1, Pawel Grzechnik 1
1School of Biosciences, University of Birmingham

Here we present a protocol for DNAzyme-dependent cleavage of RNA. This enables fast and site-dependent analysis of RNA 2’-O-methylation. This approach can be used for the preliminary or major assessment of snoRNA activity.

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Chemistry

Synthesis of Monodisperse Cylindrical Nanoparticles via Crystallization-driven Self-assembly of Biodegradable Block Copolymers
Zachary Coe 1, Andrew Weems 1, Andrew P. Dove 1, Rachel K. O'Reilly 1
1Department of Chemistry, University of Birmingham

Crystallization-driven self-assembly (CDSA) displays the unique ability to fabricate cylindrical nanostructures of narrow length distributions. The organocatalyzed ring-opening polymerization of ε-caprolactone and subsequent chain extensions of methyl methacrylate and N,N-dimethyl acrylamide are demonstrated. A living CDSA protocol that produces monodisperse cylinders up to 500 nm in length is outlined.

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Developmental Biology

Genotyping and Quantification of In Situ Hybridization Staining in Zebrafish
Tomasz Dobrzycki *1,2, Monika Krecsmarik *1,2, Rui Monteiro 1,2,3
1MRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, 2BHF Centre of Research Excellence, 3Institute of Cancer and Genomic Sciences, University of Birmingham

Gene editing technologies have enabled researchers to generate zebrafish mutants to investigate gene function with relative ease. Here, we provide a guide to perform parallel embryo genotyping and quantification of in situ hybridization signals in zebrafish. This unbiased approach provides greater accuracy in phenotypical analyses based on in situ hybridization.

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Behavior

Human Circadian Phenotyping and Diurnal Performance Testing in the Real World
Elise R. Facer-Childs 1,2,3, Benita Middleton 1, Andrew P. Bagshaw 2, Debra J. Skene 1
1Chronobiology, Faculty of Health & Medical Sciences, University of Surrey, 2Centre for Human Brain Health, University of Birmingham, 3Turner Institute for Brain and Mental Health, School of Psychological Sciences, Monash University

Here, we present a method to investigate diurnal rhythms in performance following accurate categorization of participants into circadian phenotype groups based on the Munich ChronoType Questionnaire, gold standard circadian phase biomarkers and actigraphic measures.

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Environment

Using Tg(Vtg1:mcherry) Zebrafish Embryos to Test the Estrogenic Effects of Endocrine Disrupting Compounds
Zsolt Csenki 1, Ákos Horváth 1, Illés Bock 1, Edina Garai 1, Flóra Kerekes 1, Erna Vásárhelyi 1, Balázs Kovács 1, Béla Urbányi 1, Ferenc Mueller 2, Katalin Bakos 1
1Department of Aquaculture, Institute of Aquaculture and Environmental Safety, Faculty of Agricultural and Environmental Sciences, Szent István University, 2Institute of Cancer and Genomic Sciences, College of Medical and Dental Sciences, University of Birmingham

Present here is a detailed protocol for the use of zebrafish embryos Tg(vtg1: mCherry) for the detection of estrogenic effects. The protocol covers the propagation of the fish and treatment of embryos, and emphasizes the detection, documentation, and the evaluation of fluorescent signals induced by endocrine disrupting compounds (EDC).

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Bioengineering

The Quantification of Injectability by Mechanical Testing
Thomas E. Robinson 1, Erik A. B. Hughes 1, Neil M. Eisenstein 1, Liam M. Grover 1, Sophie C. Cox 1
1School of Chemical Engineering, University of Birmingham

Presented here is a protocol for quantitatively evaluating the injectability of a material through a syringe-needle system using a standard mechanical testing rig.

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Bioengineering

Imaging and Quantification of the Area of Fast-Moving Microbubbles Using a High-Speed Camera and Image Analysis
Nina Vyas 1, Mehdi Mahmud 2, Qianxi X Wang 2, A. Damien Walmsley 1
1School of Dentistry, University of Birmingham, 2Department of Mathematics, University of Birmingham

Cavitation microbubbles are imaged using a high-speed camera attached to a zoom lens. The experimental setup is explained, and image analysis is used to calculate the area of the cavitation. Image analysis is done using ImageJ.

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Chemistry

Nanoparticle Tracking Analysis of Gold Nanoparticles in Aqueous Media through an Inter-Laboratory Comparison
Sophie M. Briffa 1, Jo Sullivan 2, Agnieszka Siupa 2, Pauline Carnell-Morris 2, Michele Carboni 2, Kerstin Jurkschat 3, Ruud J. B. Peters 4, Carolin Schultz 5, Kang Hee Seol 6, Sook-Jin Kwon 6, Sehee Park 7, Tae Hyun Yoon 6,7, Colin Johnston 3, Stephen Lofts 8, Eugenia Valsami-Jones 1
1School of Geography, Earth and Environmental Sciences, University of Birmingham, 2Malvern Panalytical, 3Department of Materials, University of Oxford, 4Wageningen Food Safety Research, 5UK Centre for Ecology & Hydrology, 6Institute for Next Generation Material Design, Hanyang University, 7Department of Chemistry, College of Natural Sciences, Hanyang University, 8UK Centre for Ecology & Hydrology

The protocol described here aims to measure the hydrodynamic diameter of spherical nanoparticles, more specifically gold nanoparticles, in aqueous media by means of Nanoparticle Tracking Analysis (NTA). The latter involves tracking the movement of particles due to Brownian motion and implementing the Stokes-Einstein equation to obtain the hydrodynamic diameter.

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Chemistry

Capillary Electrophoresis Mass Spectrometry Approaches for Characterization of the Protein and Metabolite Corona Acquired by Nanomaterials
Andrew J. Chetwynd *1, Wei Zhang *2, Klaus Faserl *3, James A. Thorn 4, Iseult Lynch 1, Rawi Ramautar 2, Herbert H. Lindner 3
1School of Geography Earth and Environmental Sciences, University of Birmingham, 2Biomedical Microscale Analytics, Leiden University, 3Institute of Medical Biochemistry, Medical University of Innsbruck, 4AB Sciex UK Ltd

Here we present a protocol to characterize the complete biomolecular corona, proteins, and metabolites, acquired by nanomaterials from biofluids using a capillary electrophoresis – mass spectrometry approach.

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JoVE Core

UV-Vis Spectroscopic Characterization of Nanomaterials in Aqueous Media
Ana C. Quevedo 1, Emily Guggenheim 1, Sophie M. Briffa 1, Jessica Adams 2,3, Stephen Lofts 2, Minjeong Kwak 4, Tae Geol Lee 4, Colin Johnston 5, Stephan Wagner 6, Timothy R. Holbrook 6, Yves U. Hachenberger 7, Jutta Tentschert 7, Nicholas Davidson 1, Eugenia Valsami-Jones 1
1School of Geography, Earth and Environmental Sciences, University of Birmingham, 2UK Centre for Ecology and Hydrology, 3Natural England, 4Center for Nanosafety Metrology, Korea Research Institute of Standards and Science (KRISS), 5Department of Materials, University of Oxford, 6Department of Analytical Chemistry, Helmholtz-Centre for Environmental Research, 7Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR)

This study presents the benchmarking results for an interlaboratory comparison (ILC) designed to test the standard operating procedure (SOP) developed for gold (Au) colloid dispersions characterized by ultraviolet-visible Spectroscopy (UV-Vis), amongst six partners from the H2020 ACEnano project for sample preparation, measurement, and analysis of the results.

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Immunology and Infection

Antibiotic Efficacy Testing in an Ex vivo Model of Pseudomonas aeruginosa and Staphylococcus aureus Biofilms in the Cystic Fibrosis Lung
Niamh E. Harrington *1, Esther Sweeney *1, Ilyas Alav 1,2, Freya Allen 1, John Moat 1,3, Freya Harrison 1
1School of Life Science, University of Warwick, 2Institute of Microbiology and Infection, University of Birmingham, 3Warwick Antimicrobial Screening Facility, School of Life Science, University of Warwick

This workflow can be used to perform antibiotic susceptibility testing using an established ex vivo model of bacterial biofilm in the lungs of individuals with cystic fibrosis. Use of this model could enhance the clinical validity of MBEC (minimal biofilm eradication concentration) assays.

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Biology

Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells
Nina Vyas *1, Nina Perry *1, Chidinma A. Okolo 1, Ilias Kounatidis 1, Thomas M. Fish 1, Kamal L. Nahas 1,2, Archana Jadhav 1, Mohamed A. Koronfel 1, Johannes Groen 3, Eva Pereiro 3, Ian M. Dobbie 4, Maria Harkiolaki 1
1Harwell Science and Innovation Campus, Beamline B24, Diamond Light Source, 2Division of Virology, Department of Pathology, University of Cambridge, 3ALBA Synchrotron, Beamline 09 - MISTRAL, 4Micron Advanced Imaging Consortium, Department of Biochemistry, University of Oxford

This protocol demonstrates how to image biological cryo-preserved samples using cryo-structured illumination microscopy. We demonstrate the methodology by imaging the cytoskeleton of U2OS cells.

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Genetics

Sample Preparation to Bioinformatics Analysis of DNA Methylation: Association Strategy for Obesity and Related Trait Studies
Natália Yumi Noronha *1, Guilherme da Silva Rodrigues *1, Marcela Augusta de Souza Pinhel 1, Jean-Baptiste Cazier 2,3, Lígia Moriguchi Watanabe 1, Albert Nobre Menezes 4, Carlos Roberto Bueno 5, Carolina Ferreira Nicoletti 1, Bruno Affonso Parenti de Oliveira 1, Isabelle Mello Schineider 6, Isabella Harumi Yonehara Noma 7, Igor Caetano Dias Alcarás 8, Fernando Barbosa 9, Carla Barbosa Nonino 6
1Department of Internal Medicine, Ribeirão Preto Medical School, University of São Paulo, 2Centre for Computational Biology, University of Birmingham, 3Institute of Cancer and Genomic Sciences, College of Medical and Dental Sciences, University of Birmingham, 4Cancer Genetics and Evolution Laboratory, Cancer Research UK, Institute of Genetics & Molecular Medicine, The University of Edinburgh, 5Ribeirão Preto School of Physical Education and Sport, University of São Paulo, 6Health Sciences Department, Ribeirao Preto Medical School, University of Sao Paulo, 7Faculty of Pharmaceutical Sciences, University of São Paulo, 8Department of Genetics, Ribeirão Preto Medical School, University of São Paulo, 9Department of Clinical, Bromatological and Toxicological Analysis, Faculty of Pharmaceutical Sciences of Ribeirao Preto, University of São Paulo

The present study describes the workflow to manage DNA methylation data obtained by microarray technologies. The protocol demonstrates steps from sample preparation to data analysis. All procedures are described in detail, and the video shows the significant steps.

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Cancer Research

A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma
Marco Y. W. Zaki *1,2, Shishir Shetty *2, Alex L. Wilkinson 2, Daniel A. Patten 2, Fiona Oakley *3, Helen Reeves *4,5
1Department of Biochemistry, Faculty of Pharmacy, Minia University, 2National Institute for Health Research Birmingham Liver Biomedical Research Unit and Centre for Liver and Gastrointestinal Research, Institute of Immunology and Immunotherapy, University of Birmingham, 3Newcastle Fibrosis Research Group, Biosciences Institute, Faculty of Medical Sciences, Newcastle University, 4Newcastle University Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, 5The Liver Unit, Department of Medicine, Freeman Hospital, Newcastle-upon-Tyne Hospitals NHS Foundation Trust

Comprehensive in vitro models that faithfully recapitulate the relevant human disease are lacking. The current study presents three-dimensional (3D) tumor spheroid creation and culture, a reliable in vitro tool to study the tumor-stromal interaction in human hepatocellular carcinoma.

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Biochemistry

Kinetx: A Combined Flow Cytometry Assay and Analysis Software Framework to Quantitatively Measure and Categorize Platelet Activation in Real-time.
Joanne L. Dunster *1, Jo L. Mitchell *1,2, Yasmin M. M. Mohammed 1, Kirk A. Taylor 1, Jonathan M. Gibbins 1, Chris I. Jones 1
1ICMR, School of Biological Sciences, University of Reading, 2Institute of Cardiovascular Sciences, University of Birmingham

Platelets react rapidly to a range of stimuli. This paper describes a real-time flow cytometry-based platelet function assay and a newly developed bespoke open-source software (Kinetx) to enable quantitative kinetic measurements of platelet granule release, fibrinogen binding, and intracellular calcium flux.

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Cancer Research

Exploring Mitochondrial Energy Metabolism of Single 3D Microtissue Spheroids Using Extracellular Flux Analysis
N. J. Coltman 1, G. Rochford 1, N. J. Hodges 1, H. Ali-Boucetta 2, J. P. Barlow 3
1School of Biosciences, University of Birmingham, 2Nanomedicine, Drug Delivery & Nanotoxicology Lab, School of Pharmacy, University of Birmingham, 3Mitochondrial Profiling Centre, University of Birmingham

These protocols will help users probe mitochondrial energy metabolism in 3D cancer cell-line-derived spheroids using Seahorse extracellular flux analysis.

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Bioengineering

Agarose Fluid Gels Formed by Shear Processing During Gelation for Suspended 3D Bioprinting
Jessica J. Senior *1, Richard J. A. Moakes *2, Megan E. Cooke 3, Samuel R. Moxon 4, Alan M. Smith 1, Liam M. Grover 2
1Department of Pharmacy, University of Huddersfield, 2School of Chemical Engineering, University of Birmingham, 3Chemical and Biological Engineering, University of Colorado Boulder, 4School of Biological Sciences, University of Manchester

Shear processing during hydrogel formation results in the production of microgel suspensions that shear-thin but rapidly restructure following the removal of shear forces. Such materials have been used as a supporting matrix for bioprinting complex, cell-laden structures. Here, methods used to manufacture the supporting bed and compatible bioinks are described.

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Biology

Assessment of Chemical Toxicity in Adult Drosophila Melanogaster
Jessica M. Holsopple *1,2, Shannon R. Smoot *1, Ellen M. Popodi 1,2, John K. Colbourne 3, Joseph R. Shaw 4, Brian Oliver 5, Thomas C. Kaufman 1, Jason M. Tennessen 1
1Department of Biology, Indiana University, 2Bloomington Drosophila Stock Center, Department of Biology, Indiana University, 3School of Biosciences, University of Birmingham, 4O'Neill School of Public and Environmental Affairs, Indiana University, 5Section of Developmental Genomics, Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Kidney and Digestive Diseases, National Institutes of Health

This protocol describes an efficient and inexpensive method that uses liquid media to assess the effects of chemical toxicants on the viability of adult Drosophila melanogaster.

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Biology

Microarray Polymer Profiling (MAPP) for High-Throughput Glycan Analysis
Cassie R. Bakshani 1,2, Jiraporn Sangta 3, Sarana Sommano 3, William G. T. Willats 1
1Department of Biology, School of Natural and Environmental Sciences, Newcastle University, 2Institute of Microbiology and Infection, College of Medical and Dental Sciences, University of Birmingham, 3Department of Plant and Soil Sciences, Faculty of Agriculture, Chiang Mai University

Microarray polymer profiling (MAPP) is a high-throughput technique for compositional analysis of glycans in biological samples.

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Biology

Evaluating Toxicity of Chemicals using a Zebrafish Vibration Startle Response Screening System
Gaëlle Hayot 1, Daniel Marcato 1,2, Christina A. Cramer von Clausbruch 1, Giuseppina Pace 1, Uwe Strähle 1,3, John K. Colbourne 4, Christian Pylatiuk 5, Ravindra Peravali 1, Carsten Weiss 1, Stefan Scholz 6, Thomas Dickmeis 1
1Institute of Biological and Chemical Systems - Biological Information Processing, Karlsruhe Institute of Technology - Campus Nord, 2DITABIS AG - Digital Biomedical Imaging Systems AG, 3Centre for Organismal Studies, Heidelberg University, 4School of Biosciences, University of Birmingham, 5Institute for Automation and Applied Informatics, Karlsruhe Institute of Technology - Campus Nord, 6Department of Bioanalytical Ecotoxicology, Helmholtz-Centre for Environmental Research - UFZ

We describe a screening system's workflow and data analysis for evaluating chemical compound toxicity based on the zebrafish embryo vibration startle response. The system records the movements of zebrafish embryos upon exposure to a vibration stimulus and allows for an integrated evaluation of general toxicity/lethality and neuromuscular toxicity.

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Medicine

Establishing a Severe Corneal Inflammation Model in Rats Based on Corneal Epithelium Curettage Combined with Corneal Sutures
Xiaoyu Tian *1,2,3, Meng Zhang *1,2, Yiming Wu 4, Liying Zhang 1,2, Lingli Zhang 1,2, Xueer Zheng 1,2, Shangkun Ou 1,2, Hao Gu 1,2
1Department of Ophthalmology, the Affiliated Hospital of Guizhou Medical University, 2School of Clinical Medicine, Guizhou Medical University, 3The Second People's Hospital of Guiyang City, 4Department of Biomedical Sciences, School of Infection, Inflammation and Immunology, College of Medicine and Health, University of Birmingham

We developed a rat model of severe corneal inflammation through corneal epithelium curettage combined with corneal sutures. The study evaluated corneal inflammation patterns, epithelial proliferation, and changes in limbal stem cells under inflammatory conditions.

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Medicine

Establishment of A Mouse Model of Aqueous Deficiency Dry Eye
Meng Zhang *1,2, Xiaoyu Tian *1,2, Yiming Wu 3, Liying Zhang 1,2, Lingli Zhang 1,2, Xueer Zheng 1,2, Shangkun Ou 1,2, Hao Gu 1,2
1Department of Ophthalmology, the Affiliated Hospital of Guizhou Medical University, 2School of Clinical Medicine, Guizhou Medical University, 3Department of Biomedical Sciences, School of Infection, Inflammation and Immunology, College of Medicine and Health, University of Birmingham

Here, we present a method to establish a mouse model of aqueous-deficient dry eye by excising the extraorbital and infraorbital lacrimal glands and evaluate the changes in the ocular surface in aqueous deficiency dry eye.

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