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5 ARTICLES PUBLISHED IN JoVE

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Bioengineering

A Zebrafish Embryo Model for In Vivo Visualization and Intravital Analysis of Biomaterial-associated Staphylococcus aureus Infection
Xiaolin Zhang 1,2, Leonie de Boer 1, Oliver W. Stockhammer 1, Dirk W. Grijpma 2,3, Herman P. Spaink 4, Sebastian A.J. Zaat 1
1Department of Medical Microbiology, Amsterdam UMC, 2Technical Medical Center, Department of Biomaterials Science and Technology, University of Twente, 3Department of Biomedical Engineering, W.J. Kolff Institute, University Medical Center Groningen, University of Groningen, 4Institute of Biology, Leiden University

The present study describes a zebrafish embryo model for in vivo visualization and intravital analysis of biomaterial-associated infection over time based on fluorescence microscopy. This model is a promising system complementing mammalian animal models such as mouse models for studying biomaterial-associated infections in vivo.

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Biology

Isolating Myofibrils from Skeletal Muscle Biopsies and Determining Contractile Function with a Nano-Newton Resolution Force Transducer
Martijn van de Locht 1, Josine M. de Winter 1, Dilson E. Rassier 2, Michiel H.B. Helmes 1,3, Coen A.C. Ottenheijm 1
1Department of Physiology, Amsterdam UMC, 2Department of Kinesiology and Physical Education, Faculty of Education, McGill University, 3IONOptix BV

Presented here is a protocol to assess the contractile properties of striated muscle myofibrils with nano-Newton resolution. The protocol employs a setup with an interferometry-based, optical force probe. This setup generates data with a high signal-to-noise ratio and enables the assessment of the contractile kinetics of myofibrils.

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JoVE Core

Acetylcholine Re-Challenge After Intracoronary Nitroglycerine Administration
Rutger G. T. Feenstra 1, Tim P. van de Hoef 1, Marcel A. M. Beijk 1, Jan J. Piek 1
1Heart Center, Department of Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam UMC

This protocol presents the acetylcholine rechallenge after nitroglycerine as an add-on procedure to spasm provocation testing. The purpose of this technique is to unmask co-existing microvascular spasm in patients with epicardial spasm and to assess the protective efficacy of nitroglycerine on a per-patient level to guide medical therapy.

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Biology

High-Throughput Contractile Measurements of Hydrogel-Embedded Intact Mouse Muscle Fibers Using an Optics-Based System
Leander A. Vonk *1, Osman Esen *1, Michaela Yuen 1,4, Tyler J. Kirby 1,2,3
1Department of Physiology, Amsterdam UMC, 2Amsterdam Cardiovascular Sciences, Heart Failures and Arrhythmias, Amsterdam UMC, 3Amsterdam Movement Sciences, Tissue Function and Regeneration, Amsterdam UMC, 4Discipline of Child and Adolescent Health, Faculty of Health and Medicine, University of Sydney

Skeletal muscle function can be assessed by quantifying the contractility of isolated muscle fibers, traditionally using laborious, low-throughput approaches. Here, we describe an optics-based, high-throughput method to quantify the contractility of hydrogel-embedded muscle fibers. This approach has applications for drug screening and therapeutic development.

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Developmental Biology

Human Ovarian Surface Epithelium Organoids as a Platform to Study Tissue Regeneration
Julieta S. Del Valle 1,2, Azra Husetic *1, Dina Diek *1, Laurens F. Rutgers *1, Joyce D. Asseler 3,4,5, Jeroen Metzemaekers 6, Norah M. van Mello 3,4,5, Susana M. Chuva de Sousa Lopes 1,2,7
1Department of Anatomy and Embryology, Leiden University Medical Center, 2The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, 3Department of Obstetrics and Gynaecology, Amsterdam University Medical Center, 4Centre of Expertise on Gender Dysphoria, Amsterdam UMC, 5Amsterdam Reproduction and Development Research Institute, 6Department of Gynaecology, Leiden University Medical Center, 7Ghent-Fertility and Stem Cell Team (G-FAST), Department of Reproductive Medicine, Ghent University Hospital

This protocol describes establishing three-dimensional (3D) tissue organoids from primary human ovarian surface epithelium (hOSE) cells. The protocol includes isolation of hOSE from freshly collected ovaries, cellular expansion of the hOSE, cryopreservation-thawing procedures, and organoid derivation. Immunofluorescence, quantitative analysis, and showcasing utility as a screening platform are included.

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