VU University, Amsterdam View Institution's Website 10 articles published in JoVE Medicine Determining and Controlling External Power Output During Regular Handrim Wheelchair Propulsion Rick de Klerk1, Riemer J. K. Vegter1, Marika T. Leving1, Sonja de Groot1,2, DirkJan H. E. J. Veeger3,4, Lucas H. V. van der Woude1,5 1Center for Human Movement Sciences, University Medical Center Groningen, 2Amsterdam Rehabilitation Research Center Reade, Amsterdam, 3Research Institute MOVE, Faculty of Behavioral and Human Movement Sciences, VU University, 43M, Delft University of Technology, Delft, 5Center for Rehabilitation, University Medical Center Groningen Accurate and standardized assessment of external power output is crucial in the evaluation of physiological, biomechanical, and perceived stress, strain, and capacity in manual wheelchair propulsion. The current article presents various methods to determine and control power output during wheelchair propulsion studies in the laboratory and beyond. Biology In Vitro Reconstitution of Light-harvesting Complexes of Plants and Green Algae Alberto Natali1, Laura M. Roy1, Roberta Croce1 1Department of Physics and Astronomy, VU University Amsterdam This protocol details the reconstitution of light-harvesting complexes in vitro. These integral membrane proteins coordinate chlorophylls and carotenoids and are responsible for harvesting light in higher plants and green algae. Biology Effect of Male Accessory Gland Products on Egg Laying in Gastropod Molluscs Sander van Iersel1, Elferra M. Swart1, Yumi Nakadera1, Nico M. van Straalen1, Joris M. Koene1 1Department of Ecological Science, Faculty of Earth and Life Sciences, VU University This video protocol demonstrates a method to study effects of seminal fluid in gastropods, using the hermaphroditic freshwater snail Lymnaea stagnalis. Neuroscience Juxtasomal Biocytin Labeling to Study the Structure-function Relationship of Individual Cortical Neurons Rajeevan T. Narayanan*1, Hemanth Mohan*1, Robin Broersen*1, Roel de Haan1, Anton W. Pieneman1, Christiaan P.J. de Kock1 1Department of Integrative Neurophysiology, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, VU University Amsterdam To understand the structure of neuronal networks, functional and morphological characterization of individual neurons is a necessity. Here, we demonstrate juxtasomal biocytin labeling, which allows electrophysiological recordings in the extracellular configuration, yet maintaining the ability to intracellularly label the neuron for post hoc reconstruction of dendritic and axonal architecture. Medicine Measuring Cardiac Autonomic Nervous System (ANS) Activity in Children Aimée E. van Dijk*1,2, René van Lien*3,4, Manon van Eijsden2,5, Reinoud J. B. J. Gemke6, Tanja G. M. Vrijkotte1, Eco J. de Geus3,4 1Department of Public Health, Academic Medical Center - University of Amsterdam, 2Department of Epidemiology, Documentation and Health Promotion, Public Health Service of Amsterdam (GGD), 3Department of Biological Psychology, VU University, 4EMGO+ Institute, VU University Medical Center, 5Institute of Health Sciences, VU University, 6Department of Pediatrics, VU University Medical Center Measurement of autonomic nervous system activity usually confines the researcher and participant to the laboratory, which may provide an intimidating environment to children. The VU University Ambulatory Monitoring System (VU-AMS) device can record cardiac autonomic control in any setting. The VU-AMS proved very amenable to testing in children. Neuroscience C. elegans Tracking and Behavioral Measurement Jirapat Likitlersuang1, Greg Stephens2,3, Konstantine Palanski1, William S. Ryu1,4 1Donnelly Centre, University of Toronto, 2Department of Physics and Astronomy, Vrije Universiteit, 3Okinawa Institute of Science and Technology, 4Department of Physics, University of Toronto We have developed a video-rate tracking microscope system that can record and quantify C. elegans behavior at high resolution and high speeds. We have also developed computational methods to reduce the dimensionality of the worm images to a fundamental set of measurements that completely describe the shape of the worm. Immunology and Infection Depletion and Reconstitution of Macrophages in Mice Shelley B. Weisser1, Nico van Rooijen2, Laura M. Sly3 1Department of Graduate Studies, University of British Columbia, 2Department of Molecular Biology, Vrije Universiteit Amsterdam, 3Department of Pediatrics, University of British Columbia Macrophages play a central role in homeostasis and pathology in many tissues. The protocol presented here describes methods for depleting macrophages in vivo, deriving polarized macrophages from bone marrow aspirates, and adoptively transferring macrophages into mice. These techniques allow determination of the role that polarized macrophages play in health and disease. Immunology and Infection Infection of Zebrafish Embryos with Intracellular Bacterial Pathogens Erica L. Benard1, Astrid M. van der Sar2, Felix Ellett3, Graham J. Lieschke3, Herman P. Spaink1, Annemarie H. Meijer1 1Department of Molecular Cell Biology, Institute of Biology, Leiden University, 2Department of Medical Microbiology and Infection Control, VU University Medical Center, 3Australian Regenerative Medicine Institute, Monash University Transparent zebrafish embryos have proved useful model hosts to visualize and functionally study interactions between innate immune cells and intracellular bacterial pathogens, such as Salmonella typhimurium and Mycobacterium marinum. Micro-injection of bacteria and multi-color fluorescence imaging are essential techniques involved in the application of zebrafish embryo infection models. Medicine High Content Screening in Neurodegenerative Diseases Shushant Jain1, Ronald E. van Kesteren2, Peter Heutink1 1Department of Clinical Genetics, VU University Medical Center, 2Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets. Neuroscience Functional Calcium Imaging in Developing Cortical Networks Julia Dawitz1, Tim Kroon1, J.J. Johannes Hjorth1, Rhiannon M. Meredith1 1Department of Integrative Neurophysiology, VU University, Amsterdam Spontaneous activity of developing neuronal networks can be measured using AM-ester forms of calcium-sensitive indicator dyes. Changes in intracellular calcium, indicating neuronal activation, are detected as transient changes in indicator fluorescence with one- or two-photon imaging. This protocol can be adapted for a range of developmentally-dependent neuronal networks in vitro.