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Rensselaer Polytechnic Institute

16 ARTICLES PUBLISHED IN JoVE

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Biology

A Technique for Serial Collection of Cerebrospinal Fluid from the Cisterna Magna in Mouse
Li Liu 1, Karen Duff 1
1Department of Pathology, Columbia University

Transgenic (Tg) mouse models of AD provide an excellent opportunity to investigate how and why Aβ or tau levels in CSF change as the disease progresses in human patients. Here, we demonstrate a refined cisterna magna puncture technique for serial CSF sampling from the mouse.

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Biology

Using an EEG-Based Brain-Computer Interface for Virtual Cursor Movement with BCI2000
J. Adam Wilson 1, Gerwin Schalk 2, Léo M. Walton 1, Justin C. Williams 1
1Department of Biomedical Engineering, University of Wisconsin-Madison, 2Wadsworth Center, New York State Dept. of Health

In this video, we demonstrate the steps required to run a brain-computer interface experiment, including setting up the EEG cap, calibrating the system, and training the user to move a cursor in two dimensions using imagined movements.

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Neuroscience

Recording Human Electrocorticographic (ECoG) Signals for Neuroscientific Research and Real-time Functional Cortical Mapping
N. Jeremy Hill 1, Disha Gupta 1,2, Peter Brunner 1,2, Aysegul Gunduz 1,2, Matthew A. Adamo 3, Anthony Ritaccio 2, Gerwin Schalk 1,2,4,5,6,7
1Wadsworth Center, New York State Department of Health, 2Department of Neurology, Albany Medical College, 3Department of Neurosurgery, Albany Medical College, 4Department of Neurosurgery, Washington University, 5Department of Biomed. Eng., Rensselaer Polytechnic Institute, 6Department of Biomed. Sci., State University of New York at Albany, 7Department of Elec. and Comp. Eng., University of Texas at El Paso

We present a method for collecting electrocorticographic signals for research purposes from humans who are undergoing invasive epilepsy monitoring. We show how to use the BCI2000 software platform for data collection, signal processing and stimulus presentation. Specifically, we demonstrate SIGFRIED, a BCI2000-based tool for real-time functional brain mapping.

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JoVE Journal

Small and Wide Angle X-Ray Scattering Studies of Biological Macromolecules in Solution
Li Liu 1, Lauren Boldon 1, Melissa Urquhart 1, Xiangyu Wang 1
1Department of Mechanical, Aerospace, and Nuclear Engineering, Rensselaer Polytechnic Institute

The demonstration of the small and wide angle X-ray scattering (SWAXS) procedure has become instrumental in the study of biological macromolecules. Through the use of the instrumentation and procedures of specific angle methods and preparation, the experimental data from the SWAXS displays the atomic and nano-scale characterization of macromolecules.

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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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Biology

Combining Magnetic Sorting of Mother Cells and Fluctuation Tests to Analyze Genome Instability During Mitotic Cell Aging in Saccharomyces cerevisiae
Melissa N. Patterson 1, Patrick H. Maxwell 1
1Department of Biological Sciences, Rensselaer Polytechnic Institute

Mutation rates in young Saccharomyces cerevisiae cells measured through fluctuation tests are used to predict mutation frequencies for mother cells of different replicative ages. Magnetic sorting and flow cytometry are then used to measure actual mutation frequencies and age of mother cells to identify any deviations from predicted mutation frequencies.

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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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Biology

Determination of Sialic Acids in Liver and Milk Samples of Wild-type and CMAH Knock-out Mice.
Cui Cao 1, Wen J. Wang 1, Ying Y. Huang 1, Hong L. Yao 1, Louis P. Conway 1, Li Liu 1, Josef Voglmeir 1
1Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University

We describe a HPLC-based method for the determination of N-acetylneuraminic acid and N-glycolylenuraminic acid in mouse liver and milk.

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Bioengineering

Fabrication of a Multiplexed Artificial Cellular MicroEnvironment Array
Yasumasa Mashimo 1,2, Momoko Yoshioka 1, Yumie Tokunaga 1, Christopher Fockenberg 1, Shiho Terada 1, Yoshie Koyama 1, Teiko Shibata-Seki 2, Koki Yoshimoto 1, Risako Sakai 1, Hayase Hakariya 1, Li Liu 1, Toshihiro Akaike 3, Eiry Kobatake 2, Siew-Eng How 4, Motonari Uesugi 1,5, Yong Chen 1,6, Ken-ichiro Kamei 1
1Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, 2Department of Life Science and Technology, School of Life Science and Technology, Tokyo Institute of Technology, 3Biomaterials Center for Regenerative Medical Engineering, Foundation for Advancement of International Science, 4Faculty of Science and Natural Resources, Universiti Malaysia Sabah, 5Institute for Chemical Research, Kyoto University, 6Ecole Normale Supérieure

This article describes the detailed methodology to prepare a Multiplexed Artificial Cellular MicroEnvironment (MACME) array for high-throughput manipulation of physical and chemical cues mimicking in vivo cellular microenvironments and to identify the optimal cellular environment for human pluripotent stem cells (hPSCs) with single-cell profiling.

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Biochemistry

Analysis of N-glycans from Raphanus sativus Cultivars Using PNGase H+
Ya-Min Du 1, Shen-Li Zheng 1, Li Liu 1, Josef Voglmeir 1, Gabriel Yedid 2
1Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, 2College of Life Science, Nanjing Agricultural University

We describe a simple and rapid method for the preparation and analysis of N-glycans from different cultivars of radish (Raphanus sativus).

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Bioengineering

A Converging Strategy for the Generation of a Virtually Sequenced cDNA Library from Unreferenced Pacific Oysters
Yong M. Lyu 1, Yu Q. Li 1, Hui B. Song 1, Juan Guo 1, Ting Wang 1, Li Liu 1, Gabriel Yedid 2, Josef Voglmeir 1
1Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University, 2College of Life Science, Nanjing Agricultural University

We describe a strategy for how to use RNA samples from unreferenced Pacific oyster specimens, and evaluate the genetic material by comparison with publicly available genome data to generate a virtually sequenced cDNA library.

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Biology

Detection of Glycosaminoglycans by Polyacrylamide Gel Electrophoresis and Silver Staining
Wells B. LaRiviere 1,2, Xiaorui Han 3,4, Kaori Oshima 1, Sarah A. McMurtry 1, Robert J. Linhardt 3, Eric P. Schmidt 1,5
1Department of Medicine, University of Colorado Anschutz Medical Campus, 2Medical Scientist Training Program, University of Colorado Anschutz Medical Campus, 3Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, 4Department of Health Sciences, Curtin University, 5Department of Medicine, Denver Health Medical Center

This report describes techniques to isolate and purify sulfated glycosaminoglycans (GAGs) from biological samples and a polyacrylamide gel electrophoresis approach to approximate their size. GAGs contribute to tissue structure and influence signaling processes via electrostatic interaction with proteins. GAG polymer length contributes to their binding affinity for cognate ligands.

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Medicine

Interphase Fluorescence in situ Hybridization of Bone Marrow Smears of Multiple Myeloma
Yalan Yu *1, Hui Shen *1, Li Liu 1, Ping Luo 1, Sanyun Wu 1, Jing He 1,2, Xiqin Tong 1, Yufeng Shang 1, Liang Shao 1, Fuling Zhou 1
1Department of Hematology, Zhongnan Hospital of Wuhan University, 2Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology

Here, we present a protocol for improving the success of interphase fluorescence in situ hybridization detection on bone marrow smears from multiple myeloma patients.

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Bioengineering

A Micropatterning Assay for Measuring Cell Chirality
Haokang Zhang 1,2, Kacey Ronaldson-Bouchard 3, Gordana Vunjak-Novakovic 3,4, Leo Q. Wan 1,2,5,6
1Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 2Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, 3Department of Biomedical Engineering, Columbia University, 4Department of Medicine, Columbia University, 5Department of Biological Sciences, Rensselaer Polytechnic Institute, 6Center for Modeling, Simulation, and Imaging in Medicine, Rensselaer Polytechnic Institute

We present a protocol for determining multicellular chirality in vitro, using the micropatterning technique. This assay allows for automatic quantification of the left-right biases of various types of cells and can be used for screening purposes.

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Biology

Directly Measuring Forces Within Reconstituted Active Microtubule Bundles
Jacob Palumbo *1, Ellinor Tai *1, Scott Forth 1
1Department of Biological Sciences and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute

Here, we present a protocol for reconstituting microtubule bundles in vitro and directly quantifying the forces exerted within them using simultaneous optical trapping and total internal reflection fluorescence microscopy. This assay allows for nanoscale-level measurement of the forces and displacements generated by protein ensembles within active microtubule networks.

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Cancer Research

Non-Destructive Evaluation of Regional Cell Density Within Tumor Aggregates Following Drug Treatment
Cassandra L. Roberge 1, Ling Wang 2, Margarida Barroso 2, David T. Corr 1
1Rensselaer Polytechnic Institute, 2Albany Medical College

The present protocol develops an image-based technique for rapid, non-destructive, and label-free regional cell density and viability measurement within 3D tumor aggregates. Findings revealed a cell-density gradient, with higher cell densities in core regions than outer layers in developing aggregates and predominantly peripheral cell death in HER2+ aggregates treated with Trastuzumab.

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