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10 ARTICLES PUBLISHED IN JoVE

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Biology

Ovariectomy and 17β-estradiol Replacement in Rats and Mice: A Visual Demonstration
Jakob O. Ström 1, Annette Theodorsson 1, Edvin Ingberg 1, Ida-Maria Isaksson 1, Elvar Theodorsson 1
1Clinical Chemistry and Neurosurgery, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University

Ovariectomy and subsequent replacement of 17β-estradiol by means of silastic capsules and peroral Nutella are demonstrated in rats and mice.

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Immunology and Infection

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection
Clara Braian 1, Mattias Svensson 2, Susanna Brighenti 2, Maria Lerm 1, Venkata R. Parasa 1,2
1Department of Clinical and Experimental Medicine, Linköping University, 2Department of Medicine, Karolinska Institute

Human tuberculosis infection is a complex process, which is difficult to model in vitro. Here we describe a novel 3D human lung tissue model that recapitulates the dynamics that occur during infection, including the migration of immune cells and early granuloma formation in a physiological environment.

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Neuroscience

Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes
Maryam Bagheri 1, Arjang Rezakhani 1, Mehrdad Roghani 2, Mohammad T. Joghataei 3,4, Simin Mohseni 1
1Department of Clinical and Experimental Medicine, Linköping University, 2Neurophysiology Research Center, Shahed University, 3Cellular and Molecular Research Center, Iran University of Medical Sciences, 4School of Advanced Technologies in Medicine, Iran University of Medical Sciences

Astrocytes in the CNS change their functional and structural properties in response to harmful stimuli. This report presents a protocol for assessment of three-dimensional astrocyte morphology in diseased conditions or after therapeutic interventions.

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Behavior

A Method for Evaluating the Reinforcing Properties of Ethanol in Rats without Water Deprivation, Saccharin Fading or Extended Access Training
Eric Augier 1, Russell S. Dulman 2, Erick Singley 2, Markus Heilig 1
1Center for Social and Affective Neuroscience, IKE, Linköping University, 2Laboratory of Clinical and Translational Studies, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health

This protocol describes a novel and efficient method to quickly initiate operant responding for ethanol in rats that, contrary to standard methods, does not require water deprivation or saccharin/sucrose fading to initiate responding.

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Neuroscience

Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
Anna Castells-Nobau *1, Bonnie Nijhof *1, Ilse Eidhof 1, Louis Wolf 2, Jolanda M. Scheffer-de Gooyert 1, Ignacio Monedero 3,4, Laura Torroja 3, Jeroen A.W.M. van der Laak 2,5, Annette Schenck *1
1Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 2Microscopical Imaging Centre (MIC), Radboud University Medical Center, 3Department of Biology, Universidad Autónoma de Madrid, 4Department of Clinical and Experimental Medicine, Linköping University, 5Department of Pathology, Radboud University Medical Center

Two image analysis algorithms, "Drosophila NMJ Morphometrics" and "Drosophila NMJ Bouton Morphometrics" were created, to automatically quantify nine morphological features of the Drosophila neuromuscular junction (NMJ).

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Biochemistry

Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging
Sofie Nyström 1, Marcus Bäck 1, K. Peter R. Nilsson 1, Per Hammarström 1
1IFM-Department of Chemistry, Linköping University

Amyloid deposition is a hallmark of different diseases and afflicts many different organs. This paper describes the application of luminescent conjugated oligothiophene fluorescence staining in combination with fluorescence microscopy techniques. This staining method represents a powerful tool for detection and exploration of protein aggregates in both clinical and scientific setups.

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Neuroscience

Diffuse Reflectance Spectroscopy: Getting the Capillary Refill Test Under One's Thumb
Joakim Henricson 1,2, Rani Toll John 1,3, Chris D. Anderson 4,5, Daniel Björk Wilhelms 1,2
1Department of Emergency Medicine, Local Health Care Services in Central Östergötland, Region Östergötland, 2Division of Drug Research, Department of Medical and Health Sciences, Faculty of Health Sciences, Linköping University, 3Division of Neuro and Inflammation Science, Department of Clinical and Experimental Medicine, Linköping University, 4Division of Cell Biology, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, 5Department of Dermatology and Venerology, Heart and Medicine Center, Region Östergötland

This protocol describes how the use of diffuse polarization spectroscopy can improve the clinical usefulness of the capillary refill test. We suggest a more detailed analysis of the course of the capillary refill in healthy volunteers using diffuse reflectance spectroscopy videos and new informatic endpoints.

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JoVE Core

Using Facial Electromyography to Assess Facial Muscle Reactions to Experienced and Observed Affective Touch in Humans
Anbjørn Ree 1, India Morrison 2, Håkan Olausson 2, Uta Sailer 1, Markus Heilig 2, Leah M. Mayo 2
1Department of Behavioral Sciences in Medicine, Institute of Basic Medical Sciences, Faculty of Medicine, University of Oslo, 2Center for Social and Affective Neuroscience, Department of Clinical and Experimental Medicine, Linköping University

We describe a protocol to assess facial muscle activity in response to experienced and observed tactile stimulation using facial electromyography.

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Biochemistry

Kinetic Screening of Nuclease Activity using Nucleic Acid Probes
Alien Balian *1,2,3, Javier Garcia Gonzalez *1,2,3, Nora Bastida 1,3, Khadija-Tul Kubra Akhtar 1,3, Baris A. Borsa 1,2,3, Frank J. Hernandez 1,2,3
1Department of Physics, Chemistry and Biology, Linköping University, 2Wallenberg Centre for Molecular Medicine (WCMM), 3Nucleic Acids Technologies Laboratory (NAT-lab), Linköping University

Altered nuclease activity has been associated with different human conditions, underlying its potential as a biomarker. The modular and easy to implement screening methodology presented in this paper allows the selection of specific nucleic acid probes for harnessing nuclease activity as a biomarker of disease.

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Immunology and Infection

Systematic Scoring Analysis for Intestinal Inflammation in a Murine Dextran Sodium Sulfate-Induced Colitis Model
Vicky Garcia-Hernandez 1, Philipp-Alexander Neumann 2, Stefan Koch 3,4, Renae Lyons 1, Asma Nusrat 1, Charles A. Parkos 1
1Department of Pathology, University of Michigan, 2Department of Surgery, Klinikum Rechts der Isar, Technische Universität München, 3Wallenberg Centre for Molecular Medicine, Linköping University, 4Department of Biomedical and Clinical Sciences (BKV), Linköping University

Systematic scoring of intestinal inflammation using a free computer-assisted system is a powerful tool to quantitatively compare histopathological changes in colitis models characterized by the presence of ulcers and inflammatory changes. Histological colitis score evaluation strengthens clinical observations and facilitates data interpretation.

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