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University of Milano-Bicocca

6 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions
Achille Broggi 1,2, Clara Cigni 1, Ivan Zanoni 1,2,3, Francesca Granucci 1,3
1Department of Biotechnology and Biosciences, University of Milano-Bicocca, 2Boston Children's Hospital, Division of Gastroenterology, Harvard Medical School, 3Humanitas Clinical and Research Center

The skin is home to a complex immune cell network. We describe an efficient methodology for the digestion of mouse skin, from different parts of the animal's body, in order to obtain a single-cell suspension and analyze the different leukocyte populations resident in the skin by flow cytometry.

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Developmental Biology

Induction of Acute Skeletal Muscle Regeneration by Cardiotoxin Injection
Ombretta Guardiola 1, Gennaro Andolfi 1, Mario Tirone 2,3, Francescopaolo Iavarone 1, Silvia Brunelli 2, Gabriella Minchiotti 1
1Stem Cell Fate Laboratory, Institute of Genetics and Biophysics "A. Buzzati Traverso", CNR, 2School of Medicine and Surgery, University of Milano-Bicocca, 3Division of Immunology, Transplantation and Infectious Diseases, San Raffaele Scientific Institute

This manuscript describes a detailed protocol to induce acute skeletal muscle regeneration in adult mice and subsequent manipulations of the muscles, such as dissection, freezing, cutting, routine staining, and myofiber cross-sectional area analysis.

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Immunology and Infection

Deep Dermal Injection As a Model of Candida albicans Skin Infection for Histological Analyses
William Santus 1, Francesca Mingozzi 1, Marina Vai 1, Francesca Granucci *1, Ivan Zanoni *1,2
1Department of Biotechnology and Biosciences, University of Milano-Bicocca, 2Harvard Medical School and Division of Gastroenterology, Boston Children's Hospital

Here we describe a protocol that allows histological and molecular analysis of skin samples after Candida albicans intradermal injection. This protocol maintains the structural integrity of the skin and allows for the localization of tissue-resident or newly recruited immune cells as well as the pathogen distribution.

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Neuroscience

Real-time Video Projection in an MRI for Characterization of Neural Correlates Associated with Mirror Therapy for Phantom Limb Pain
Faddi G. Saleh Velez 1,2, Camila B. Pinto 1,3, Emma S. Bailin 4, Marionna Münger 1, Andrew Ellison 5, Beatriz T. Costa 1, David Crandell 6, Nadia Bolognini 7,8, Lotfi B. Merabet 4, Felipe Fregni 1
1Laboratory of Neuromodulation & Center for Clinical Research Learning, Department of Physical Medicine and Rehabilitation, Harvard Medical School, Spaulding Rehabilitation Hospital, 2University of Chicago Medical Center, Department of Neurology, University of Chicago, 3Department of Neuroscience and Behavior, Psychology Institute, University of Sao Paulo, 4The Laboratory for Visual Neuroplasticity, Department of Ophthalmology, Massachusetts Eye and Ear Infirmary, Harvard Medical School, 5Center for Biomedical Imaging, Department of Anatomy and Neurobiology, Boston University School of Medicine, 6Spaulding Rehabilitation Hospital, Harvard Medical School, 7Department of Psychology & Milan Center for Neuroscience, University of Milano-Bicocca, 8Neuropsychological Laboratory, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Istituto Auxologico Italiano

We present a novel combined behavioral and neuroimaging protocol employing real-time video projection for the purpose of characterizing the neural correlates associated with mirror therapy within the magnetic resonance imaging scanner environment in leg amputee subjects with phantom limb pain.

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JoVE Journal

Exploiting Live Imaging to Track Nuclei During Myoblast Differentiation and Fusion
Giorgia Careccia *1,2, Federica Colombo *1,3, Mario Tirone 1, Alessandra Agresti 1, Marco E. Bianchi 1,2, Samuel Zambrano *1,2, Emilie Vénéreau *1
1Chromatin Dynamics Unit, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) San Raffaele Scientific Institute, 2Vita-Salute San Raffaele University, 3Department of Electronics, Information and Bioengineering, Politecnico di Milano

Skeletal muscle differentiation is a highly dynamic process, which particularly relies on nuclear positioning. Here, we describe a method to track nuclei movements by live cell imaging during myoblast differentiation and myotube formation and to perform a quantitative characterization of nuclei dynamics by extracting information from automatic tracking.

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Medicine

Monitoring Lung Function with Electrical Impedance Tomography in the Intensive Care Unit
Glasiele C. Alcala *1, Gamonmas Ekkapat *1,2, Kyle J. Medeiros 1, Caio C. A Morais 1, Yi Xin 1, Valentina Giammatteo 1,3, Giovanni Bruno 1, Alice Nova 1,4, Hatus Wanderley 1, Talisa Bühl 1, Marcus Victor 1,6, Timothy G. Gaulton 1, Carolyn J. La Vita 5, Marcelo B. P. Amato 7, Maurizio Ceradda 1, Lorenzo Berra 1, Roberta Ribeiro De Santis Santiago 1
1Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Harvard Medical School, 2Department of Anesthesiology, Faculty of Medicine, Chulalongkorn University. King Chulalongkorn Memorial Hospital, 3Department of Anesthesiology and Intensive Care Medicine, Catholic University of Sacred Heart, 4School of Medicine and Surgery, University of Milano-Bicocca, 5Department of Respiratory Care, Massachusetts General Hospital, Harvard Medical School, 6Electronics Engineering, Aeronautics Institute of Technology, 7Instituto do Coracao (InCor), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo

Electrical Impedance Tomography is a non-invasive, radiation-free, real-time pulmonary ventilation monitoring tool. By measuring impedance changes in the thorax, it can visualize the distribution of air on a breath-by-breath basis. Initially intended for ventilation monitoring, electrical impedance tomography can also measure perfusion via intravenous injection of a saline solution.

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