University of Milano-Bicocca

6 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions

Achille Broggi^1,2, Clara Cigni¹, 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.

Developmental Biology

Induction of Acute Skeletal Muscle Regeneration by Cardiotoxin Injection

Ombretta Guardiola¹, Gennaro Andolfi¹, Mario Tirone^2,3, Francescopaolo Iavarone¹, Silvia Brunelli², Gabriella Minchiotti¹

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.

Immunology and Infection

Deep Dermal Injection As a Model of Candida albicans Skin Infection for Histological Analyses

William Santus¹, Francesca Mingozzi¹, Marina Vai¹, 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.

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. Bailin⁴, Marionna Münger¹, Andrew Ellison⁵, Beatriz T. Costa¹, David Crandell⁶, Nadia Bolognini^7,8, Lotfi B. Merabet⁴, Felipe Fregni¹

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.

JoVE Journal

Exploiting Live Imaging to Track Nuclei During Myoblast Differentiation and Fusion

Giorgia Careccia^*1,2, Federica Colombo^*1,3, Mario Tirone¹, Alessandra Agresti¹, 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.

Medicine

Monitoring Lung Function with Electrical Impedance Tomography in the Intensive Care Unit

Glasiele C. Alcala^*1, Gamonmas Ekkapat^*1,2, Kyle J. Medeiros¹, Caio C. A Morais¹, Yi Xin¹, Valentina Giammatteo^1,3, Giovanni Bruno¹, Alice Nova^1,4, Hatus Wanderley¹, Talisa Bühl¹, Marcus Victor^1,6, Timothy G. Gaulton¹, Carolyn J. La Vita⁵, Marcelo B. P. Amato⁷, Maurizio Ceradda¹, Lorenzo Berra¹, Roberta Ribeiro De Santis Santiago¹

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.