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18 ARTICLES PUBLISHED IN JoVE

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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

Fluorescent Nanoparticles for the Measurement of Ion Concentration in Biological Systems
J. Matthew Dubach 1, Mary K. Balaconis 1, Heather A. Clark 2
1Bioengineering Department, Northeastern University, 2Department of Pharmaceutical Sciences, Northeastern University

Fluorescent nanoparticles produced in our lab are used for imaging ion concentrations and ion fluxes in biological systems such as cells during signaling and interstitial fluid during physiological homeostasis.

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Bioengineering

Therapeutic Gene Delivery and Transfection in Human Pancreatic Cancer Cells using Epidermal Growth Factor Receptor-targeted Gelatin Nanoparticles
Jing Xu 1, Mansoor Amiji 1
1Department of Pharmaceutical Sciences, School of Pharmacy, Northeastern University

Type B gelatin-based engineered nanovectors system (GENS) was developed for systemic gene delivery and transfection in the treatment of pancreatic cancer. By modification with epidermal growth factor receptor (EGFR) specific peptide on the surface of nanparticles, they could target on EGFR receptor and release plasmid under reducing environment, such as high intracellular glutathione concentrations.

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Neuroscience

Designing and Implementing Nervous System Simulations on LEGO Robots
Daniel Blustein 1, Nikolai Rosenthal 2, Joseph Ayers 1
1Marine Science Center, Northeastern University, 2Fachrichtung Bionik, Bremen University of Applied Sciences

An approach to neural network modeling on the LEGO Mindstorms robotics platform is presented. The method provides a simulation tool for invertebrate neuroscience research in both the research lab and the classroom. This technique enables the investigation of biomimetic robot control principles.

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Engineering

Sputter Growth and Characterization of Metamagnetic B2-ordered FeRh Epilayers
Chantal Le Graët 1, Mark A. de Vries 1,2, Mathew McLaren 1,3, Richard M.D. Brydson 3, Melissa Loving 4, Don Heiman 5, Laura H. Lewis 4, Christopher H. Marrows 1
1School of Physics and Astronomy, University of Leeds, 2Institute of Materials Research, University of Leeds, 3School of Chemistry, University of Edinburgh, 4Department of Chemical Engineering, Northeastern University, 5Department of Physics, Northeastern University

A method to prepare epitaxial layers of ordered alloys by sputtering is described. The B2-ordered FeRh compound is used as an example, as it displays a metamagnetic transition that depends sensitively on the degree of chemical order and the exact composition of the alloy.

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Bioengineering

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology
Xiaofeng Cui *1,2,3, Guifang Gao *2,4, Tomo Yonezawa 5,6, Guohao Dai 1
1Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 2Stemorgan Inc., 3Institute of Advanced Study, Technical University of Munich, 4Institute of Virology, School of Medicine, Wuhan University, 5Department of Molecular and Experimental Medicine, The Scripps Research Institute, 6Research Institute for Biomedical Sciences, Tokyo University of Science

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

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Developmental Biology

Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors
Wei Wen 1, Jian-Ping Zhang 1, Wanqiu Chen 2, Cameron Arakaki 2, Xiaolan Li 1,2, David Baylink 2, Gary D. Botimer 3, Jing Xu 1, Weiping Yuan 1, Tao Cheng 1,4,5,6,7, Xiao-Bing Zhang 1,2
1State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 2Division of Regenerative Medicine, Department of Medicine, Loma Linda University, 3Department of Orthopaedic Surgery, Loma Linda University, 4Center for Stem Cell Medicine, Chinese Academy of Medical Sciences, 5Department of Stem Cell & Regenerative Medicine, Peking Union Medical College, 6Collaborative Innovation Center for Cancer Medicine, 7Tianjin Key Laboratory of Blood Cell Therapy and Technology

This protocol describes a detailed method for efficient generation of integration-free iPSCs from human adult peripheral blood cells. With the use of four oriP/EBNA-based episomal vectors to express the reprogramming factors, KLF4, MYC, BCL-XL, or OCT4 and SOX2, thousands of iPSC colonies can be obtained from 1 mL of peripheral blood.

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JoVE Journal

Light-mediated Reversible Modulation of the Mitogen-activated Protein Kinase Pathway during Cell Differentiation and Xenopus Embryonic Development
Vishnu V. Krishnamurthy *1, Aurora J. Turgeon *2, John S. Khamo 1, Payel Mondal 1, Savanna R. Sharum 1, Wenyan Mei 2, Jing Yang 2, Kai Zhang 1,3,4
1Department of Biochemistry, University of Illinois at Urbana-Champaign, 2Department of Comparative Biosciences, University of Illinois at Urbana-Champaign, 3Neuroscience Program, University of Illinois at Urbana-Champaign, 4Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign

This protocol describes an optogenetic strategy to modulate mitogen-activated protein kinase (MAPK) activity during cell differentiation and Xenopus embryonic development. This method allows for the reversible activation of the MAPK signaling pathway in mammalian cell culture and in multicellular live organisms, like Xenopus embryos, with high spatial and temporal resolution.

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Bioengineering

Patterning Bioactive Proteins or Peptides on Hydrogel Using Photochemistry for Biological Applications
Taylor B. Dorsey 1,2,3, Alexander Grath 1,2, Cancan Xu 4, Yi Hong 4, Guohao Dai 1,2,3
1Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 2Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 3Department of Bioengineering, Northeastern University, 4Department of Bioengineering, University of Texas at Arlington

In this method, we use photopolymerization and click chemistry techniques to create protein or peptide patterns on the surface of polyethylene glycol (PEG) hydrogels, providing immobilized bioactive signals for the study of cellular responses in vitro.

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Bioengineering

Synthesis of Functionalized 10-nm Polymer-coated Gold Particles for Endothelium Targeting and Drug Delivery
Ming J. Cheng 1, Priya Prabakaran 1, Rajiv Kumar 2,3, Srinivas Sridhar 1,2,3, Eno E. Ebong 1,4,5
1Department of Chemical Engineering, Northeastern University, 2Nanomedicine Science and Technology Center, Northeastern University, 3Department of Physics, Northeastern University, 4Departments of Bioengineering, Northeastern University, 5Department of Neuroscience, Albert Einstein College of Medicine

We describe a method of synthesizing biocompatible 10-nm gold nanoparticles, functionalized by coating poly-ethylene glycol onto the surface. These particles can be used in vitro and in vivo for delivering therapeutics to nanoscale cellular and extracellular spaces that are difficult to access with conventional nanoparticle sizes.

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Chemistry

Elemental-sensitive Detection of the Chemistry in Batteries through Soft X-ray Absorption Spectroscopy and Resonant Inelastic X-ray Scattering
Jinpeng Wu 1,2, Shawn Sallis 2,3, Ruimin Qiao 2, Qinghao Li 2,4, Zengqing Zhuo 2,5, Kehua Dai 2,6, Zixuan Guo 2,7, Wanli Yang 2
1Geballe Laboratory for Advanced Materials, Stanford University, 2Advanced Light Source, Lawrence Berkeley National Laboratory, 3Department of Materials Science and Engineering, Binghamton University, 4School of Physics, National Key Laboratory of Crystal Materials, Shandong University, 5School of Advanced Materials, Peking University Shenzhen Graduate School, 6School of Metallurgy, Northeastern University, 7Department of Chemical Engineering, University of California-Santa Barbara

Here, we present a protocol for typical experiments of soft X-ray absorption spectroscopy (sXAS) and resonant inelastic X-ray scattering (RIXS) with applications in battery material studies.

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Biochemistry

Polysome Profiling in Leishmania, Human Cells and Mouse Testis
Zemfira N. Karamysheva 2, Elena B. Tikhonova 1, Petar N. Grozdanov 1, James C. Huffman 2,3, Kristen R. Baca 1,3, Alexander Karamyshev 1, R. Brian Denison 1, Clinton C. MacDonald 1, Kai Zhang 2, Andrey L. Karamyshev 1
1Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, 2Department of Biological Sciences, Texas Tech University, 3CISER (Center for the Integration of STEM Education & Research), Texas Tech University

The overall goal of polysome profiling technique is analysis of translational activity of individual mRNAs or transcriptome mRNAs during protein synthesis. The method is important for studies of protein synthesis regulation, translation activation and repression in health and multiple human diseases.

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Engineering

Low-cost Custom Fabrication and Mode-locked Operation of an All-normal-dispersion Femtosecond Fiber Laser for Multiphoton Microscopy
Kai Zhang 1,2, Nima Davoudzadeh 1,2, Guillaume Ducourthial 1,2, Bryan Q. Spring 1,2,3
1Translational Biophotonics Cluster, Northeastern University, 2Department of Physics, Northeastern University, 3Department of Bioengineering, Northeastern University

A method is presented to build a custom low-cost, mode-locked femtosecond fiber laser for potential applications in multiphoton microscopy, endoscopy, and photomedicine. This laser is built using commercially available parts and basic splicing techniques.

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Bioengineering

Characterization of Intra-Cartilage Transport Properties of Cationic Peptide Carriers
Armin Vedadghavami 1, Shikhar Mehta 1, Ambika G. Bajpayee 1,2
1Department of Bioengineering, Northeastern University, 2Department of Mechanical & Industrial Engineering, Northeastern University

This protocol determines equilibrium uptake, depth of penetration and non-equilibrium diffusion rate for cationic peptide carriers in cartilage. Characterization of transport properties is critical for ensuring an effective biological response. These methods can be applied for designing an optimally charged drug carriers for targeting negatively charged tissues.

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Biology

Single-Cell Proteomics Preparation for Mass Spectrometry Analysis Using Freeze-Heat Lysis and an Isobaric Carrier
Aleksandra A. Petelski *1, Nikolai Slavov 1,2,3, Harrison Specht *1
1Department of Bioengineering, Northeastern University, 2Barnett Institute, Northeastern University, 3Department of Biology, Northeastern University

In this protocol, we describe how to prepare mammalian cells for single-cell proteomics analysis via mass spectrometry using commercially available reagents and equipment, with options for both manual and automatic pipetting.

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Bioengineering

Synthesis of Graphene-Hydroxyapatite Nanocomposites for Potential Use in Bone Tissue Engineering
Sougata Ghosh 1,2,3, Tanay Bhagwat 3, Rohini Kitture 4, Sirikanjana Thongmee 1, Thomas J. Webster 3,5,6,7
1Department of Physics, Faculty of Science, Kasetsart University, 2Department of Microbiology, School of Science, RK. University, 3Department of Chemical Engineering, Northeastern University, 4Defence Institute of Advanced Technology Campus, Navyukti Innovations Pvt. Ltd., 5Department of Biomedical Engineering, Hebei University of Technology, 6Department of Chemistry, Saveetha School of Engineering, Institute of Medical and Technical Science, Saveetha University, 7UFPI - Universidade Federal do Piauí

Novel nanocomposites of graphene nanoribbons and hydroxyapatite nanoparticles were prepared using solution-phase synthesis. These hybrids when employed in bioactive scaffolds can exhibit potential applications in tissue engineering and bone regeneration.

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Immunology and Infection

Deciphering the Molecular Mechanism and Function of Pore-Forming Toxins Using Leishmania major
Chaitanya S. Haram 1, Samrat Moitra 1, Rilee Keane 1, Elana Breslav 1, Kai Zhang 1, Peter A. Keyel 1
1Department of Biological Sciences, Texas Tech University

Presented here is a protocol using Leishmania major promastigotes to determine the binding, cytotoxicity, and signaling induced by pore-forming toxins. A proof-of-concept with streptolysin O is provided. Other toxins can also be used to leverage the genetic mutants available in L. major to define new mechanisms of toxin resistance.

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Biology

Assembly and Operation of a Cooling Stage to Immobilize C. elegans on Their Culture Plates
Yao L. Wang 1, Noa W. F. Grooms 1, Claire W. Ma 1, Samuel H. Chung 1
1Department of Bioengineering, Northeastern University

This paper describes protocols for constructing and operating a cooling stage to immobilize C. elegans on their original cultivation plates en masse.

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