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Technical University Munich

11 ARTICLES PUBLISHED IN JoVE

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Biology

Born Normalization for Fluorescence Optical Projection Tomography for Whole Heart Imaging
Claudio Vinegoni 1,2, Daniel Razansky 3, Jose-Luiz Figueiredo 1,2, Lyuba Fexon 1,2, Misha Pivovarov 1,2, Matthias Nahrendorf 1,2, Vasilis Ntziachristos 3, Ralph Weissleder 1,2
1Center for Systems Biology, Harvard Medical School, 2Center for Systems Biology, MGH - Massachusetts General Hospital, 3Institute for Biological and Medical Imaging, Technical University of Munich and Helmholtz Center Munich

We suggest a Born normalized approach for Optical Projection Tomography (BnOPT) that accounts for the absorption properties of imaged samples to obtain accurate and quantitative fluorescence tomographic reconstructions. We use the proposed algorithm to reconstruct the fluorescence molecular probe distribution within small animal organs.

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Biology

Mesoscopic Fluorescence Tomography for In-vivo Imaging of Developing Drosophila
Claudio Vinegoni 1, Daniel Razansky 2, Chrysoula Pitsouli 3, Norbert Perrimon 3, Vasilis Ntziachristos 2, Ralph Weissleder 1
1Center for Systems Biology, Massachusetts General Hospital, 2Institute for Biological and Medical Imaging (IBMI), Technical University of Munich and Helmholtz Center Munich, 3Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute

Mesoscopic fluorescence tomography operates beyond the penetration limits of tissue-sectioning fluorescence microscopy. The technique is based on multi-projection illumination and a photon transport description. We demonstrate in-vivo whole-body 3D visualization of the morphogenesis of GFP-expressing wing imaginal discs in Drosophila melanogaster.

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Medicine

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology
Lucia M.A. Crane 1, George Themelis 2, K. Tim Buddingh 1, Niels J. Harlaar 1, Rick G. Pleijhuis 1, Athanasios Sarantopoulos 2, Ate G.J. van der Zee 3, Vasilis Ntziachristos 2, Gooitzen M. van Dam 1
1Department of Surgery, Division of Surgical Oncology, University Medical Center Groningen, 2Helmholtz Zentrum, Technical University Munich, 3Department of Obstetrics and Gynaecology, University Medical Center Groningen

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

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Medicine

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis
Marcella A. Calfon 1, Amir Rosenthal 1,2, Georgios Mallas 1,3, Adam Mauskapf 1, R. Nika Nudelman 2, Vasilis Ntziachristos 2, Farouc A. Jaffer 1
1Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital, Harvard Medical School, 2Institute for Biological and Medical Imaging, Helmholtz Zentrum München und Technische Universität München, 3Department of Electrical and Computer Engineering, Northeastern University

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

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Bioengineering

Wideband Optical Detector of Ultrasound for Medical Imaging Applications
Amir Rosenthal 1, Stephan Kellnberger 1, Murad Omar 1, Daniel Razansky 1, Vasilis Ntziachristos 1
1Institute for Biological and Medical Imaging (IBMI), Technical University of Munich and Helmholtz Center Munich

Optical detection of ultrasound is impractical in many imaging scenarios because it often requires stable environmental conditions. We demonstrate an optical technique for ultrasound sensing in volatile environments with miniaturization and sensitivity levels appropriate for optoacoustic imaging in restrictive scenarios, e.g. intravascular applications.

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Medicine

Tissue-simulating Phantoms for Assessing Potential Near-infrared Fluorescence Imaging Applications in Breast Cancer Surgery
Rick Pleijhuis 1, Arwin Timmermans 1, Johannes De Jong 1, Esther De Boer 1, Vasilis Ntziachristos 2, Gooitzen Van Dam 1
1Department of Surgery, University Medical Center Groningen, 2Helmholtz Zentrum Munich, Technical University of Munich

Near-infrared fluorescence (NIRF) imaging may improve therapeutic outcome of breast cancer surgery by enabling intraoperative tumor localization and evaluation of surgical margin status. Using tissue-simulating breast phantoms containing fluorescent tumor-simulating inclusions, potential clinical applications of NIRF imaging in breast cancer patients can be assessed for standardization and training purposes.

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Developmental Biology

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues
Bartosz Ditkowski 1, Tiago R Veloso 1, Martyna Bezulska-Ditkowska 1,2, Andreas Lubig 3, Stefan Jockenhoevel 3, Petra Mela 3, Ramadan Jashari 4, Marc Gewillig 1, Bart Meyns 5, Marc F Hoylaerts 2, Ruth Heying 1
1Cardiovascular Developmental Biology, Department of Cardiovascular Sciences, KU Leuven, 2Centre for Molecular and Vascular Biology, Department of Cardiovascular Sciences, KU Leuven, 3Department of Biohybrid & Medical Textiles, AME - Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, 4European Homograft Bank, Saint Jean Clinique, 5Division of Clinical Cardiac Surgery, Department of Cardiovascular Sciences, KU Leuven

We describe an in-house designed in vitro flow chamber model, which allows the investigation of bacterial adherence to graft tissues.

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Genetics

Optimized High Quality DNA Extraction from Formalin-Fixed Paraffin-Embedded Human Atherosclerotic Lesions
Sabine Bauer 1,2,4, Jessica Pauli 1,2, Jaroslav Pelisek 1,3
1Department for Vascular and Endovascular Surgery, Klinikum rechts der Isar, Technical University Munich, 2partner site Munich Heart Alliance, Deutsches Zentrum für Herz-Kreislaufforschung (DZHK), 3Department of Vascular Surgery, University Hospital Zurich, 4Department of Experimental Cardiology, German Heart Centre, Technical University Munich

Here, we present a protocol for semi-automated DNA extraction from formalin-fixed paraffin-embedded lesions of human carotid arteries. The tissue lysis is performed without toxic xylene, which is followed by an automated DNA extraction protocol, including a second lysis step, binding of DNA to paramagnetic particles for cellulose based binding, washing steps, and DNA elution.

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Medicine

Development and Evaluation of 3D-Printed Cardiovascular Phantoms for Interventional Planning and Training
Maximilian Grab 1,2, Carina Hopfner 3, Alena Gesenhues 4, Fabian König 1,2, Nikolaus A. Haas 3, Christian Hagl 1, Adrian Curta 4, Nikolaus Thierfelder 1
1Department of Cardiac Surgery, Ludwig Maximilian University Munich, 2Chair of Medical Materials and Implants, Technical University of Munich, 3Department Pediatric Cardiology and Pediatric Intensive Care, Ludwig Maximilian University Munich, 4Department of Radiology, Ludwig Maximilian University Munich

Here we present development of a mock circulation setup for multimodal therapy evaluation, pre-interventional planning, and physician-training on cardiovascular anatomies. With the application of patient-specific tomographic scans, this setup is ideal for therapeutic approaches, training, and education in individualized medicine.

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Biochemistry

Mass-Sensitive Particle Tracking to Characterize Membrane-Associated Macromolecule Dynamics
Frederik Steiert 1,2, Tamara Heermann 1, Nikolas Hundt 3, Petra Schwille 1
1Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, 2Department of Physics, Technical University Munich, 3Department of Cellular Physiology, Biomedical Center (BMC), Ludwig-Maximilians-Universität München

This protocol describes an iSCAT-based image processing and single-particle tracking approach that enables the simultaneous investigation of the molecular mass and the diffusive behavior of macromolecules interacting with lipid membranes. Step-by-step instructions for sample preparation, mass-to-contrast conversion, movie acquisition, and post-processing are provided alongside directions to prevent potential pitfalls.

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Medicine

Combining 3D-Printing and Electrospinning to Manufacture Biomimetic Heart Valve Leaflets
Benedikt Freystetter 1, Maximilian Grab 1,2, Linda Grefen 1, Lara Bischof 1, Lorenz Isert 3, Petra Mela 2, Deon Bezuidenhout 4, Christian Hagl 1,5, Nikolaus Thierfelder 1
1Department of Cardiac Surgery, Ludwig Maximilians University Munich, 2Chair of Medical Materials and Implants, Technical University Munich, 3Faculty for Chemistry and Pharmacy, Ludwig Maximilians University Munich, 4Cardiovascular Research Unit, University of Cape Town, 5DZHK (German Centre for Cardiovascular Research), partner site Munich Heart Alliance

The presented method offers an innovative way for engineering biomimetic fiber structures in three-dimensional (3D) scaffolds (e.g., heart valve leaflets). 3D-printed, conductive geometries were used to determine shape and dimensions. Fiber orientation and characteristics were individually adjustable for each layer. Multiple samples could be manufactured in one setup.

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