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University of Southern Denmark

18 ARTICLES PUBLISHED IN JoVE

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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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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

Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat
Morten Hox 1, Travis Mann-Gow 2, Lars Lund 3,4, Peter Zvara 1,4
1Biomedical Laboratory and the Research Unit of Urology, Department of Clinical Research, University of Southern Denmark, 2Department of Surgery, University of Vermont, 3Research Unit of Urology, Department of Clinical Research, University of Southern Denmark, 4Department of Urology, Odense University Hospital

This study describes a simplified surgical procedure and technique for performing cavernous nerve stimulation with the isolation of the nerve-electrode complex using silicone glue and intracavernous pressure measurement.

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JoVE Journal

Assessing Collagen and Elastin Pressure-dependent Microarchitectures in Live, Human Resistance Arteries by Label-free Fluorescence Microscopy
Maria Bloksgaard 1, Bjarne Thorsted 2, Jonathan R. Brewer 2, Jo G. R. De Mey 1,3
1Department of Cardiovascular and Renal Research, Institute of Molecular Medicine, University of Southern Denmark, 2Department of Biochemistry and Molecular Biology, University of Southern Denmark, 3Department of Cardiac, Thoracic and Vascular Surgery, Odense University Hospital

We describe simultaneous mechanical testing and 3D-imaging of the arterial wall of isolated, live human resistance arteries, and Fiji and Ilastik image analyses for the quantification of elastin and collagen spatial organization and volume densities. We discuss the use of these data in mathematical models of arterial wall mechanics.

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Immunology and Infection

Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples
Maiken W. Rosenstierne 1, Christopher E. Jensen 1, Anders Fomsgaard 1,2
1Virus Research & Development Laboratory, Statens Serum Institut, 2Infectious Disease Research Unit, University of Southern Denmark

Here, we present a protocol for the rapid virus nucleic acid extraction from the virus-inactivated whole blood. The extraction is performed directly in the blood collection tubes and requires no equipment or electricity. The method is not dependent on laboratory facilities and can be used anywhere (e.g., in field hospitals).

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Biochemistry

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli
Patrick Rosendahl Andreassen 1, Jens Sivkær Pettersen 1, Mikkel Jørgensen 1
1Department of Biochemistry and Molecular Biology, University of Southern Denmark

Site-directed mutagenesis is a technique used to introduce specific mutations in deoxyribonucleic acid (DNA). This protocol describes how to do site-directed mutagenesis with a 2-step and 3-step polymerase chain reaction (PCR) based approach, which is applicable to any DNA fragment of interest.

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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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Medicine

Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy
Sabrina T. Hansen 1,2, Charlotte H. Jensen 2,3,4, Jens A. Sørensen 2,4,5, Søren P. Sheikh 2,3,4, Lars Lund 1,2,4
1Department of Urology, Odense University Hospital, 2Clinical Institute, University of Southern Denmark, 3Laboratory of Molecular and Cellular Cardiology, Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, 4Center for Vascular Regeneration, Odense University Hospital, 5Research Unit for Plastic Surgery, Odense University Hospital

Precise disclosure of methods and protocols is crucial for large scale uptake of stem cell therapies. Here, we present a protocol to isolate adipose-derived regenerative cells, used for a single intracavernous injection as treatment of erectile dysfunction (ED) following radical prostatectomy (RP).

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Immunology and Infection

Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes
Jeff Granhøj 1, Henrik Dimke 1,2, Per Svenningsen 1
1Department of Molecular Medicine, University of Southern Denmark, 2Department of Nephrology, Odense University Hospital

Here we present a method to select for novel variants of the E. coli biotin-protein ligase BirA that biotinylates a specific target peptide. The protocol describes the construction of a plasmid for the bacterial display of the target peptide, generation of a BirA library, selection and characterization of BirA variants.

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Medicine

A Revised Method for Inducing Secondary Lymphedema in the Hindlimb of Mice
Alexander Wiinholt 1,2, Mads G. Jørgensen 2, Amar Bučan 1,2, Farima Dalaei 1,2, Jens A. Sørensen 2
1University of Southern Denmark, 2Department of Plastic Surgery, Odense University Hospital

This animal model enables researchers to induce statistically significant secondary lymphedema in the hindlimb of mice, lasting at least 8 weeks. The model can be used to study the pathophysiology of lymphedema and to investigate novel treatment options.

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Medicine

Direct Drug Delivery to Kidney via the Renal Artery
Ulrika Dahlqvist 1, Tajana Tesan Tomic 2, Magnus Söderberg 3, Jane Stubbe 4, Camilla Enggaard 4, Anette Ericsson 5, Alex-Xianghua Zhou 1, Anna Björnson Granqvist 1, Lena William-Olsson 1
1Bioscience Renal, Research and Early Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, 2Cell/mol Pharmacology, Research and Early Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D, 3CVRM Safety, Safety, Clinical Pharmacology & Safety Sciences, R&D, 4Department of Cardiovascular and Renal Research, University of Southern Denmark, 5Clinical Renal, Late-stage Development, Cardiovascular, Renal and Metabolism, BioPharmaceuticals R&D

This manuscript describes a method for targeted delivery to a single kidney via a catheter placed in the infrarenal abdominal aorta in the mouse.

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Biochemistry

Quantification of Subcellular Glycogen Distribution in Skeletal Muscle Fibers using Transmission Electron Microscopy
Rasmus Jensen 1, Niels Ørtenblad 2, Cristiano di Benedetto 3, Klaus Qvortrup 3, Joachim Nielsen 2
1Research center for applied health science, University College South Denmark, 2Department of Sports Science and Clinical Biomechanics, University of Southern Denmark, 3Department of Biomedical Sciences, Core Facility for Integrated Microscopy, University of Copenhagen

A modified post-fixation procedure increases the contrast of glycogen particles in tissue. This paper provides a step-by-step protocol describing how to handle the tissue, conduct the imaging, and use stereological methods to obtain unbiased and quantitative data on fiber type-specific subcellular glycogen distribution in skeletal muscle.

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Biochemistry

Semi-Automated Phenotypic Analysis of Functional 3D Spheroid Cell Cultures
Stephanie Stransky 1, Dejauwne Young 1, Karoline Mikkelsen 2,3, Annemette Præstegaard Thulesen 2, Helle Sedighi Frandsen 3, Simone Sidoli 1
1Department of Biochemistry, Albert Einstein College of Medicine, 2Department of Biochemistry and Molecular Biology, University of Southern Denmark, 3CelVivo ApS

We present a protocol for growing high-reproducible spheroids and their phenotypic characterization using image capture and proteomics.

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Medicine

Radial Endobronchial Ultrasound and Electromagnetic Navigation Bronchoscopy with Fluoroscopy for the Diagnosis of Peripheral Lung Lesions
Amanda Dandanell Juul 1,2, Alexis Pulga 3, Arman Arshad 1
1Department of Respiratory Medicine, Odense University Hospital, 2Odense Respiratory Research Unit (ODIN), Department of Clinical Research, University of Southern Denmark, 3Department of Respiratory Medicine, Bispebjerg Hospital

Diagnosing small lung tumors is quite difficult using a bronchoscope alone. Electromagnetic navigation bronchoscopy is used to locate the lesion, similar to the Global Positioning System. Radial endobronchial ultrasound and fluoroscopy confirm the correct location and monitor the sampling.

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JoVE Core

Local Anesthetic Thoracoscopy for Undiagnosed Pleural Effusion
Uffe Bodtger 1,2, José M. Porcel 3, Rahul Bhatnagar 4,5, Nick Maskell 4,5, Mohammed Munavvar 6,7, Casper Jensen 1, Paul Frost Clementsen 1,8, Daniel Bech Rasmussen 1,2
1Respiratory Research Unit PLUZ, Department of Respiratory Medicine, Zealand University Hospital, 2Institute of Regional Health Research, University of Southern Denmark, 3Pleural Medicine Unit, Department of Internal Medicine, Hospital Universitari Arnau de Vilanova, IRBLleida, 4Respiratory Department, Southmead Hospital, North Bristol NHS Trust, 5Academic Respiratory Unit, University of Bristol, 6Lancashire Teaching Hospitals, 7University of Central Lancashire, 8Centre for HR and Education, Copenhagen Academy for Medical Education and Simulation

Local anesthetic thoracoscopy (LAT) is essential for diagnosing recurrent, undiagnosed pleural effusion when a guideline-based workup fails to provide a specific cause. LAT can be performed as a day-case procedure by chest physicians. Here, we present a step-by-step approach for a successful and safe procedure.

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Medicine

Using the Endoscope for Endobronchial Ultrasound in the Esophagus
Mohammad A. Issa 1, Paul F. Clementsen 2, Christian B. Laursen 3,4, Peter Vilmann 5,6, Ida S. Christiansen 7, Laurence Crombag 8, Uffe Bodtger 1,9
1Pulmonary Research Unit (PLUZ), Department of Respiratory Medicine, Zealand University Hospital, 2Copenhagen Academy for Medical Education and Simulation (CAMES), Copenhagen University Hospital, Rigshospitalet, 3Department of Respiratory Medicine, Odense University Hospital, 4Odense Respiratory Research Unit (ODIN), Department of Clinical Research, University of Southern Denmark, 5Department of Surgery, Copenhagen University Hospital, 6Department of Clinical Medicine, University of Copenhagen, 7Department of Pathology, Copenhagen University Hospital, Rigshospitalet, 8Department of Respiratory Medicine, University Medical Center, University of Amsterdam, 9Institute for Regional Health Research, University of Southern Denmark

Transesophageal ultrasound (EUS-B) is a safe and feasible procedure using the echoendobronchoscope (EBUS) in esophagus and stomach. After identifying six anatomical landmarks, additional structures can be identified and biopsied, sparing subsequent diagnostic sessions. Thus, EUS-B is an ideal continuation of bronchoscopy and EBUS in diagnosing lung cancer and other diseases.

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Medicine

Transbronchial Lung Cryobiopsy for Diagnosing Interstitial Lung Diseases and Peripheral Pulmonary Lesions - A Stepwise Approach
Jesper R. Davidsen 1,2, Christian B. Laursen 1,2, Søren H. Skaarup 3, Sissel B. Kronborg-White 3, Amanda D. Juul 1,2
1South Danish Center for Interstitial Lung Diseases (SCILS), Department of Respiratory Medicine, Odense University Hospital, 2Odense Respiratory Research Unit (ODIN), Department of Clinical Research, University of Southern Denmark, 3Department of Respiratory Diseases and Allergy, Aarhus University Hospital

Transbronchial lung cryobiopsy (TBLC) for diagnosing interstitial lung disease and peripheral pulmonary lesions is a high-yield diagnostic and safe procedure. We describe a stepwise approach to conduct TBLC for the different indications mentioned with a flexible bronchoscope, which might be helpful for novice bronchoscopists performing TBLC.

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Medicine

A Stepwise Approach for Performing Ultrasound Guided Transthoracic Lung Biopsy
Christian B. Laursen 1,2, Rahul Bhatnagar 2,3,4, Amanda Dandanell Juul 1,2
1Department of Respiratory Medicine, Odense University Hospital, 2Odense Respiratory Research Unit (ODIN), Department of Clinical Research, University of Southern Denmark, 3Respiratory Medicine Department, North Bristol NHS Trust, Southmead Hospital, 4Academic Respiratory Unit, University of Bristol

Transthoracic ultrasound-guided lung biopsy represents a safe, cost-effective, and efficient approach for patients presenting with subpleural lung lesions suspected of malignancy. Employing a systematic, step-by-step process is crucial to achieve optimal patient selection, minimize complication risks, and maximize diagnostic accuracy.

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