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Aarhus University

40 ARTICLES PUBLISHED IN JoVE

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JoVE Journal

Identification and Analysis of Myogenic Progenitors In Vivo During Acute Skeletal Muscle Injury by High-Dimensional Single-Cell Mass Cytometry
Richard Medeiros 1,2
1Department of Biomedicine, Aarhus University, 2Aarhus Institute of Advanced Studies, Aarhus University

The protocol presented here enables the identification and high-dimensional analysis of muscle stem and progenitor cells by single-cell mass cytometry and their purification by FACS for in-depth studies of their function. This approach can be applied to study regeneration dynamics in disease models and test the efficacy of pharmacological interventions.

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Medicine

Basic Surgical Techniques in the Göttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion
Kaare S. Ettrup 1,2, Andreas N. Glud 2, Dariusz Orlowski 2, Lise M. Fitting 1, Kaare Meier 1, Jens Christian Soerensen 1, Carsten R. Bjarkam 1,2, Aage K. Olsen Alstrup 3
1Department of Neurosurgery, Aarhus University Hospital, 2Department of Neurobiology, Institute of Anatomy, Faculty of Health Sciences, Aarhus University, 3Positron Emission Tomography (PET) Centre, Aarhus University Hospital

The use of domestic and miniature pigs in science has increased significantly in recent years. By demonstrating how to perform intubation, transurethral bladder catheterization, femoral artery and vein catheterization, as well as transcardial perfusion, we aim to further increase the value of Göttingen minipigs in biomedical research.

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Biology

Identification and Analysis of Mouse Erythroid Progenitors using the CD71/TER119 Flow-cytometric Assay
Miroslav Koulnis *1, Ramona Pop *1, Ermelinda Porpiglia *1, Jeffrey R. Shearstone *1, Daniel Hidalgo 1, Merav Socolovsky 1
1Department of Pediatrics and Department of Cancer Biology, University of Massachusetts Medical School

A flow-cytometric method for identification and molecular analysis of differentiation-stage-specific murine erythroid progenitors and precursors, directly in freshly –harvested mouse bone marrow, spleen or fetal liver. The assay relies on cell-surface markers CD71, Ter119, and cell size.

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Medicine

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice
Mette Richner 1, Ole J. Bjerrum 2, Anders Nykjaer 1, Christian B. Vaegter 1
1The Lundbeck Foundation Research Center MIND, Department of Biomedicine, Aarhus University, 2Department of Pharmacology and Pharmacotherapy, Faculty of Pharmaceutical Sciences, University of Copenhagen

The Spared Nerve Injury animal model is described here as a mouse model of peripheral neuropathic pain following partial denervation of the sciatic nerve by lesioning the tibial and common peroneal nerve branches, leaving the remaining sural nerve intact. Behavioral modification resulting from mechanical allodynia is quantified by von Frey filaments.

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Biology

Easy Measurement of Diffusion Coefficients of EGFP-tagged Plasma Membrane Proteins Using k-Space Image Correlation Spectroscopy
Eva C. Arnspang 1, Jennifer S. Koffman 1, Saw Marlar 1, Paul W. Wiseman 2, Lene N. Nejsum 1
1Institute of Molecular Biology and Genetics and Interdisciplinary Nanoscience Center, Aarhus University, 2Departments of Chemistry and Physics, McGill University

This paper provides a step by step guide to the fluctuation analysis technique k-Space Image Correlation Spectroscopy (kICS) for measuring diffusion coefficients of fluorescently labeled plasma membrane proteins in live mammalian cells.

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Immunology and Infection

Ratiometric Imaging of Extracellular pH in Dental Biofilms
Sebastian Schlafer 1, Irene Dige 1
1Department of Dentistry, Aarhus University

A pH-sensitive ratiometric dye is used in combination with confocal laser scanning microscopy and digital image analysis to monitor extracellular pH in dental biofilms in real-time.

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Chemistry

X-ray Powder Diffraction in Conservation Science: Towards Routine Crystal Structure Determination of Corrosion Products on Heritage Art Objects
Robert E. Dinnebier 1, Andrea Fischer 2, Gerhard Eggert 2, Tomče Runčevski 1, Nanna Wahlberg 3
1Max-Planck-Institute for Solid State Research, 2State Academy of Art and Design Stuttgart, 3Center for Materials Crystallography, Department of Chemistry and iNANO, Aarhus University

Modern high resolution X-ray powder diffraction (XRPD) in the laboratory is used as an efficient tool to determine crystal structures of long-known corrosion products on historic objects.

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Biochemistry

SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry
Jakob V. Larsen 1, Claus M. Petersen 1
1Department of Biomedicine, Aarhus University

The present protocol describes how Western blotting and immunocytochemistry combined with inhibition of lysosomal enzymes can be used to demonstrate the downregulation of Ciliary Neurotrophic Factor Receptor-α (CNTFRα), which is conveyed by the interaction between Cytokine-like Factor-1 (CLF-1) and the endocytic receptor sorLA.

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Neuroscience

Hydraulic Extrusion of the Spinal Cord and Isolation of Dorsal Root Ganglia in Rodents
Mette Richner 1, Sara B. Jager 1, Piotr Siupka 1, Christian B. Vaegter 1
1Department of Biomedicine, Aarhus University

Here, we present a protocol for hydraulic extrusion of the spinal cord as well as identification and isolation of specific dorsal root ganglia (DRGs) in the same rodent. Compared to standard spinal cord isolation methods, this method is significantly faster and reduces the risk of tissue damage.

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JoVE Core

Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal
Carsten R. Bjarkam 1, Dariusz Orlowski 2, Laura Tvilling 2, Johannes Bech 2, Andreas N. Glud 2, Jens-Christian H. Sørensen 2
1Department of Neurosurgery, Clinical Institute of Medicine, Aalborg University Hospital, 2Center of Experimental Neuroscience (Cense), Department of Neurosurgery, Institute of Clinical Medicine, Aarhus University Hospital

The goal of this paper and instructional video is to describe how to expose and remove the postmortem pig brain and pituitary gland in an intact state, suitable for subsequent macroscopic and histological analysis.

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Medicine

Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model
Travis K. Mann-Gow *1, Troy R. Larson *1, Chrissie T. Wøien 2, Thomas M. Andersen 2, Karl-Erik Andersson 3,4, Peter Zvara 1,2
1Department of Surgery, University of Vermont, 2Department of Urology and Biomedical Laboratory, University of Southern Denmark, 3Wake Forest Institute for Regenerative Medicine, Wake Forest University, 4Institute for Clinical Medicine, Aarhus University

This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization.

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Medicine

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells
Simone S. E. Nielsen 1, Piotr Siupka 1, Ana Georgian 2, Jane E. Preston 2, Andrea E. Tóth 1, Siti R. Yusof 2,3, N. Joan Abbott 2, Morten S. Nielsen 1
1Lundbeck Foundation Research Initiative on Brain Barriers and Drug Delivery, Department of Biomedicine, Aarhus University, 2Institute of Pharmaceutical Science, King's College London, 3HICoE Centre for Drug Research, Universiti Sains Malaysia

The aim of the protocol is to present an optimized procedure for the establishment of an in vitro blood-brain barrier (BBB) model based on primary porcine brain endothelial cells (pBECs). The model shows high reproducibility, high tightness, and is suitable for studies of transport and intracellular trafficking in drug discovery.

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Bioengineering

Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
Jeremy J. Flint 1,2, Kannan Menon 2,3, Brian Hansen 4, John Forder 2,3,5, Stephen J. Blackband 1,2,6
1Department of Neuroscience, University of Florida, 2McKnight Brain Institute, University of Florida, 3Department of Biomedical Engineering, University of Florida, 4Center for Functionally Integrative Neuroscience, Aarhus University, 5Department of Radiology, University of Florida, 6National High Magnetic Field Laboratory, Florida State University

The current protocol describes a method by which users can maintain viability of acute hippocampal and cortical slice preparations during the collection of magnetic resonance microscopy data.

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Neuroscience

An Improved Method for Collection of Cerebrospinal Fluid from Anesthetized Mice
Nastasia K-H Lim 1,2, Visse Moestrup 3,4, Xiao Zhang 1, Wen-An Wang 5, Arne Møller 3,4,6, Fu-De Huang 1,4
1Shanghai Advanced Research Institute, University of Chinese Academy of Sciences, Chinese Academy of Science, 2Shanghai Institute of Materia Medica, University of Chinese Academy of Sciences, Chinese Academy of Science, 3Center of Functionally Integrative Neuroscience, Department of Clinical Medicine, Aarhus University, 4Sino-Danish Center for Education and Research (SDC), 5Department of Neurology, Xinhua Hospital Chongming Branch Affiliated to Shanghai Jiao Tong University School of Medicine, 6Department of Nuclear Medicine and PET-centre, Aarhus University Hospital

This protocol describes an improved technique for the abundant collection of cerebrospinal fluid (CSF) with no contamination from blood. With greater sample collection and purity, more analyses can be performed using CSF to further our understanding of diseases that affect the brain and spinal cord.

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Genetics

Investigation of RNA Synthesis Using 5-Bromouridine Labelling and Immunoprecipitation
Rikke H. Kofoed 1, Cristine Betzer 1, Søren Lykke-Andersen 2, Ewa Molska 2, Poul H. Jensen 1
1DANDRITE - Danish Research Institute of Translational Neuroscience, Dept. of Biomedicine, Aarhus University, 2Dept. of Molecular Biology and Genetics, Aarhus University

This method can be used to measure RNA synthesis. 5-Bromouridine is added to cells and incorporated into synthesized RNA. RNA synthesis is measured by RNA extraction immediately after labelling, followed by 5-Bromouridine-targeted immunoprecipitation of labelled RNA and analysis by reverse transcription and quantitative polymerase chain reaction.

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Medicine

2D and 3D Echocardiography in the Axolotl (Ambystoma Mexicanum)
Anita Dittrich 1, Mathias Møller Thygesen 1, Henrik Lauridsen 1
1Department of Clinical Medicine, Aarhus University

Here we present echocardiography protocols for two-dimensional and three-dimensional image acquisition of the beating heart of the axolotl salamander (Ambystoma mexicanum), a model species in heart regeneration. These methods allow for longitudinal evaluation of cardiac function at a high spatiotemporal resolution.

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Cancer Research

Virus Delivery of CRISPR Guides to the Murine Prostate for Gene Alteration
Maria Riedel 1, Martin F. Berthelsen 1, Latifa Bakiri 2, Erwin F. Wagner 2, Martin K. Thomsen 1
1Department of Clinical Medicine, Aarhus University, 2GDD, Cancer Cell Biology Program, National Cancer Research Center (CNIO)

This protocol describes a newly established method for virus delivery to the murine prostate. Using either CRISPR/Cas9 technology, gene overexpression, or Cre recombinase delivery, the technique allows orthotopic alteration of gene expression and implements a novel mouse model for prostate cancer.

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Medicine

A Pulmonary Trunk Banding Model of Pressure Overload Induced Right Ventricular Hypertrophy and Failure
Stine Andersen 1, Jacob Gammelgaard Schultz 1, Sarah Holmboe 1, Julie Birkmose Axelsen 1, Mona Sahlholdt Hansen 1, Mads Dam Lyhne 1, Jens Erik Nielsen-Kudsk 1, Asger Andersen 1
1Department of Clinical Medicine and Cardiology, Aarhus University Hospital

We present a surgical method to induce right ventricular hypertrophy and failure in rats.

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Genetics

Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells
Yan Yan *1, Chi-Chih Chang *1, Morten T. Venø 1,2, Colin R. Mothershead 1, Junyi Su 1, Jørgen Kjems 1,2
1The Interdisciplinary Nanoscience Centre, Aarhus University, 2Department of Molecular Biology and Genetics, Aarhus University

This protocol demonstrates how to purify extracellular microRNAs from cell culture media for small RNA library construction and next generation sequencing. Various quality control checkpoints are described to allow readers to understand what to expect when working with low input samples like exRNAs.

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Behavior

Spotlighting Customers' Visual Attention at the Stock, Shelf and Store Levels with the 3S Model
Tobias Otterbring 1,2, Erik Wästlund 2, Poja Shams 3
1Department of Management/MAPP, Aarhus University, 2Department of Psychology & CTF, Service Research Center, Karlstad University, 3Department of Business Administration & CTF, Karlstad University

This article presents a new conceptualization of the in-store search process, the 3S Model, which captures customers’ visual attention at three distinct levels of analysis: Stock, Shelf, and Store. We illustrate the usefulness of our conceptualization through three eye-tracking studies, one from each level of analysis in the 3S Model.

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Bioengineering

Uptake of New Lipid-coated Nanoparticles Containing Falcarindiol by Human Mesenchymal Stem Cells
Emma Pipó-Ollé 1, Prasad Walke 2, Martine K. Notabi 1, Rime B. El-Houri 1, Morten Østergaard Andersen 1, David Needham 2,3,4, Eva C. Arnspang 1
1Department of Chemical Engineering, Biotechnology and Environmental Technology, University of Southern Denmark, 2Center for Single Particle Science and Engineering (SPSE), Institute for Molecular Medicine, Health Sciences, University of Southern Denmark, 3Department of Mechanical Engineering and Material Science, Duke University, 4School of Pharmacy, University of Nottingham

This article describes the encapsulation of falcarindiol in lipid-coated 74 nm nanoparticles. The cellular uptake of the nanoparticles by human stem cells into lipid droplets is monitored by fluorescent and confocal imaging. Nanoparticles are fabricated by the rapid injection method of solvent shifting, and their size is measured with the dynamic light scattering technique.

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Behavior

Transcranial Direct Current Stimulation (tDCS) of Wernicke's and Broca's Areas in Studies of Language Learning and Word Acquisition
Evgeny Blagovechtchenski 1, Daria Gnedykh 1, Diana Kurmakaeva 1, Nadezhda Mkrtychian 1, Svetlana Kostromina 1, Yury Shtyrov 1,2
1Laboratory of Behavioural Neurodynamics, St. Petersburg State University, 2Center of Functionally Integrative Neuroscience (CFIN), Department of Clinical Medicine, Aarhus University

Here, we describe a protocol for using transcranial direct current stimulation for psycho- and neurolinguistic experiments aimed at studying, in a naturalistic yet fully controlled way, the role of cortical areas of the human brain in word learning, and a comprehensive set of behavioral procedures for assessing the outcomes.

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Immunology and Infection

Interrogating Individual Autoreactive Germinal Centers by Photoactivation in a Mixed Chimeric Model of Autoimmunity
Thomas R. Wittenborn 1, Cecilia Hagert 1, Søren E Degn 1
1Department of Biomedicine, Aarhus University

This protocol describes the generation of mixed murine bone marrow chimeras with spontaneous autoimmune germinal centers, in which autoreactive lymphocytes carry a photoactivatable green fluorescent protein (PA-GFP) reporter. This provides the ability to link cellular location in tissues with downstream molecular and functional analyses.

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Chemistry

Luminescence Lifetime Imaging of O2 with a Frequency-Domain-Based Camera System
Maria Moßhammer *1, Vincent V. Scholz *2, Gerhard Holst 3, Michael Kühl 1,4, Klaus Koren 5
1Marine Biological Section, Department of Biology, University of Copenhagen, 2Center for Electromicrobiology, Aarhus University, 3PCO AG, 4Climate Change Cluster, University of Technology Sydney, 5Aarhus University Centre for Water Technology, Section for Microbiology, Department of Bioscience, Aarhus University

We describe the use of a novel, frequency-domain luminescence lifetime camera for mapping 2D O2 distributions with optical sensor foils. The camera system and image analysis procedures are described along with the preparation, calibration and application of sensor foils for visualizing the O2 microenvironment in the rhizosphere of aquatic plants.

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Immunology and Infection

Monitoring Extracellular pH in Cross-Kingdom Biofilms using Confocal Microscopy
Sebastian Schlafer 1, Mathilde Frost Kristensen 1
1Department of Dentistry and Oral Health, Aarhus University

The protocol describes the cultivation of cross-kingdom biofilms consisting of Candida albicans and Streptococcus mutans and presents a confocal microscopy-based method for the monitoring of extracellular pH inside these biofilms.

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Medicine

Contusion Spinal Cord Injury via a Microsurgical Laminectomy in the Regenerative Axolotl
Mathias Møler Thygesen 1,2, Fredrik Guldbæk-Svensson 1, Mikkel Mylius Rasmussen 2, Henrik Lauridsen 1
1Department of Clinical Medicine, Aarhus University, 2Department of Neurosurgery, Aarhus University Hospital

This manuscript presents protocols for surgically inflicting controlled blunt and sharp spinal cord injuries to a regenerative axolotl (Ambystoma mexicanum).

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Biology

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae
Chuanxian Wei *1, Yan Yan *1, Xiaoli Miao 1, Renjie Jiao 1,2,3
1Sino-French Hoffmann Institute, School of Basic Medical Sciences, Guangzhou Medical University, 2The Second Affiliated Hospital of Guangzhou Medical University, 3The State Key Laboratory of Respiratory Disease, Guangzhou Medical University

Presented here are detailed methods for the dissection and lipid droplet staining of oenocytes in Drosophila larvae using BODIPY 493/503, a lipid droplet-specific fluorescent dye.

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Immunology and Infection

Humanized NOG Mice for Intravaginal HIV Exposure and Treatment of HIV Infection
Anna H. F. Andersen 1,2, Stine S. F. Nielsen 1,2, Rikke Olesen 1,2, Katharina Mack 1, Frederik Dagnæs-Hansen 3, Niels Uldbjerg 1,4, Lars Østergaard 1,2, Ole S. Søgaard 1,2, Paul W. Denton 1,2,5, Martin Tolstrup 1,2
1Department of Clinical Medicine, Aarhus University, 2Department of Infectious Diseases, Aarhus University Hospital, 3Department of Biomedicine, Aarhus University, 4Department of Gynecology and Obstetrics, Aarhus University Hospital, 5Department of Biology, University of Nebraska at Omaha

We have developed a protocol for the generation and evaluation of a humanized and human immunodeficiency virus-infected NOG mouse model based on stem cell transplant, intravaginal human immunodeficiency virus exposure, and droplet digital PCR RNA quantification.

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Biochemistry

Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRAC
Stuart W. McKellar 1, Ivayla Ivanova 1, Robert W. van Nues 2, Ross A. Cordiner 3, Mehak Chauhan 1, Niki Christopoulou 1, Will Worboys 4, Andrew Langford 4, Torben Heick Jensen 3, Sander Granneman 1
1Centre for Engineering Biology, University of Edinburgh, 2Institute of Cell Biology, University of Edinburgh, 3Department of Molecular Biology and Genetics, Aarhus University, 4UVO3 Ltd.

Kinetic cross-linking and analysis of cDNA is a method that allows investigation of the dynamics of protein-RNA interactions in living cells at high temporal resolution. Here the protocol is described in detail, including the growth of yeast cells, UV cross-linking, harvesting, protein purification, and next generation sequencing library preparation steps.

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Developmental Biology

Cryo-injury Induced Heart Regeneration in the Axolotl and Echocardiography and Unbiased Quantitative Histology to Evaluate Regenerative Progression
Anita Dittrich 1, Henrik Lauridsen 1
1Department of Clinical Medicine (Comparative Medicine Lab), Aarhus University

Here we present a surgery protocol to induce cryoinjury to the ventricular myocardium of the axolotl. Additionally, we present a protocol to non-invasively estimate infarction fraction during the regenerative process with echocardiography and a protocol to precisely measure infarction fraction in the excised heart with unbiased quantitative histology.

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Behavior

Transition of Farm Pigs to Research Pigs using a Designated Checklist followed by Initiation of Clicker Training - a Refinement Initiative
Anders F. Thomsen *1, Birgitte S. Kousholt *1
1Department of Clinical Medicine, AUGUST Centre, Aarhus University

Refinement of porcine studies is achieved by introducing a standardized checklist and positive reinforcement training using a clicker. This work supports the collection of samples and the conduct of daily chores related to the animals.

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Behavior

A Within-Subject Experimental Design using an Object Location Task in Rats
Gülberk Bayraktar *1,2, Kristoffer Højgaard *1,3, Luc Nijssen 1,4, Tomonori Takeuchi 1,5
1Danish Research Institute of Translational Neuroscience (DANDRITE), Department of Biomedicine, Aarhus University, 2Universitätsklinikum Würzburg, Institut für Klinische Neurobiologie, 3Translational Neuropsychiatry Unit, Department of Clinical Medicine, Aarhus University, 4Faculty of Science, Radboud University, 5Center for Proteins in Memory - PROMEMO, Danish National Research Foundation, Department of Biomedicine, Aarhus University

This protocol provides detailed steps for an object location task with four repetitions using the same cohort of rats. Weak and strong encoding can produce short- and long-term memories. The flexibility of the protocol with repetition can be beneficial for studies involving surgical operations by saving time and labor.

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Medicine

Closed Chest Biventricular Pressure-Volume Loop Recordings with Admittance Catheters in a Porcine Model
Mads Dam Lyhne 1,2, Jacob Gammelgaard Schultz 1,2, Simone Juel Dragsbaek 1,2, Jacob Valentin Hansen 1,2, Christian Schmidt Mortensen 1,2, Anders Kramer 1,2, Jens Erik Nielsen-Kudsk 1,2, Asger Andersen 1,2
1Department of Cardiology Research, Aarhus University Hospital, 2Department of Clinical Medicine, Aarhus University

Here we present a closed chest approach to admittance-based bi-ventricular pressure-volume loop recordings in pigs with acute right ventricular dysfunction.

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Biology

Deep Vascular Imaging in the Eye with Flow-Enhanced Ultrasound
Christian Damsgaard 1,2, Henrik Lauridsen 3
1Aarhus Institute of Advanced Studies, Aarhus University, 2Zoophysiology, Department of Biology, Aarhus University, 3Department of Clinical Medicine, Aarhus University

We present a non-invasive ultrasound technique for generating three-dimensional angiographies in the eye without the use of contrast agents.

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Cancer Research

Simple and Fast Rolling Circle Amplification-Based Detection of Topoisomerase 1 Activity in Crude Biological Samples
Josephine Geertsen Keller 1,2, Karol Mizielinski 3, Kamilla Vandsø Petersen 1,2, Magnus Stougaard 2,3,4, Birgitta R. Knudsen 1,3, Cinzia Tesauro 3
1Department of Molecular Biology and Genetics, Aarhus University, 2Department of Clinical Medicine, Aarhus University, 3VPCIR Biosciences ApS, 4Department of Pathology, Aarhus University Hospital

A protocol for the sensitive and quantitative detection of topoisomerase 1 activity using the rolling circle enhanced enzyme activity detection assay is described. The method allows detection of topoisomerase 1 activity from purified components or cell/tissue extracts. This protocol has wide-ranging applications in any field involving detection of enzymatic activity.

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Biology

Isolation of Quiescent Stem Cell Populations from Individual Skeletal Muscles
Zofija Frimand *1, Shubhangi Das Barman *1, Troels Rønn Kjær 1, Ermelinda Porpiglia 1, Antoine de Morrée 1
1Department of Biomedicine, Aarhus University

This protocol describes the isolation of muscle stem cells and fibro-adipogenic progenitors from individual skeletal muscles in mice. The protocol involves single muscle dissection, stem cell isolation by fluorescence activated cell sorting, purity assessment by immunofluorescence staining, and quantitative measurement of S-phase entry by 5-ethynyl-2´-deoxyuridine incorporation assay.

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Medicine

Semi-Automated Planimetric Quantification of Dental Plaque Using an Intraoral Fluorescence Camera
Pernille Dukanovic Rikvold 1, Yumi Chokyu Del Rey 1, Karina Kambourakis Johnsen 1, Sebastian Schlafer 1
1Department of Dentistry and Oral Health, Section for Oral Ecology and Caries Control, Aarhus University

This study presents a semi-automated digital image analysis procedure for the planimetric quantification of disclosed dental plaque based on images acquired with an intraoral fluorescence camera. The method allows for the rapid and reliable quantification of dental plaque in the research environment.

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Biology

Identification and Analysis of Myogenic Progenitors In Vivo During Acute Skeletal Muscle Injury by High-Dimensional Single-Cell Mass Cytometry
Stig Henrik Andersen 1, Troels Rønn Kjær 1, Kedar Ghimire 1, Ermelinda Porpiglia 1,2
1Department of Biomedicine, Aarhus University, 2Aarhus Institute of Advanced Studies, Aarhus University

The protocol presented here enables the identification and high-dimensional analysis of muscle stem and progenitor cells by single-cell mass cytometry and their purification by FACS for in-depth studies of their function. This approach can be applied to study regeneration dynamics in disease models and test the efficacy of pharmacological interventions.

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Immunology and Infection

Microfluidic Approach to Resolve Simultaneous and Sequential Cytokine Secretion of Individual Polyfunctional Cells
Aline Linder *1, Kevin Portmann *1, Luca Johannes Schlotheuber 1, Alessandro Streuli 1, Wiona Sophie Glänzer 1, Klaus Eyer 1,2, Ines Lüchtefeld 1,3
1Laboratory for Functional Immune Repertoire Analysis, Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences, ETH Zürich, 2Department of Biomedicine, Aarhus University, 3Laboratory for Tumor and Stem Cell Dynamics, Institute of Molecular Health Sciences, Department of Biology, ETH Zürich

The protocol describes an advanced microfluidic platform to quantitatively measure cytokine secretion dynamics of individual human peripheral blood mononuclear cells. The platform measures up to three cytokines in parallel (IL-6, TNFα, and IL-1β) for each individual cell stimulated with lipopolysaccharide as an example.

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Medicine

A Murine Model of Pressure Overload-Induced Right Ventricular Hypertrophy and Failure by Pulmonary Trunk Banding
Anders Hammer Nielsen-Kudsk *1,2, Janne Schwab *1,2, Julie Sørensen Axelsen 1,2, Asger Andersen 1,2
1Department of Clinical Medicine, Aarhus University, 2Department of Cardiology, Aarhus University Hospital

We describe a murine model of right ventricular pressure overload-induced by pulmonary trunk banding. Detailed protocols for intubation, surgery, and phenotyping by echocardiography are included in the paper. Custom-made instruments are used for intubation and surgery, allowing for fast and inexpensive reproduction of the model.

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