Institute for Basic Science (IBS) 8 articles published in JoVE Engineering Fabrication of Micro-Patterned Chip with Controlled Thickness for High-Throughput Cryogenic Electron Microscopy Min-Ho Kang1,2, Minyoung Lee3,4, Sungsu Kang3,4, Jungwon Park3,4,5,6 1Department of Biomedical-Chemical Engineering, The Catholic University of Korea, 2Department of Biotechnology, The Catholic University of Korea, 3School of Chemical and Biological Engineering, and Institute of Chemical Processes, Seoul National University, 4Center for Nanoparticle Research, Institute of Basic Science (IBS), 5Institute of Engineering Research, College of Engineering, Seoul National University, 6Advanced Institutes of Convergence Technology, Seoul National University A newly developed micro-patterned chip with graphene oxide windows is fabricated by applying microelectromechanical system techniques, enabling efficient and high-throughput cryogenic electron microscopy imaging of various biomolecules and nanomaterials. Biochemistry Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique Yujin Kang1, Subin Bae1, Soyeong An1, Ja Yil Lee1,2 1Department of Biological Sciences, Ulsan National Institute of Science and Technology, 2Center for Genomic Integrity, Institute for Basic Science (IBS) DNA curtain, a high-throughput single-molecule imaging technique, provides a platform for real-time visualization of diverse protein-DNA interactions. The present protocol utilizes the DNA curtain technique to investigate the biological role and molecular mechanism of Abo1, a Schizosaccharomyces pombe bromodomain-containing AAA+ ATPase. Biology Extracellular Vesicle Uptake Assay via Confocal Microscope Imaging Analysis Chi-Ju Kim*1,2, Morgan D. Kuczler*1, Liang Dong1,3, Junyoung Kim2,4, Sarah R. Amend1, Yoon-Kyoung Cho2,4, Kenneth J. Pienta1 1The Brady Urological Institute, Johns Hopkins University School of Medicine, 2Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), 3Department of Urology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, 4Center for Soft and Living Matter, Institute for Basic Science (IBS) Extracellular vesicles (EVs) contribute to cellular biology and intercellular communications. There is a need for practical assays to visualize and quantify EVs uptake by the cells. The current protocol proposes the EV uptake assay by utilizing three-dimensional fluorescence imaging via confocal microscopy, following EV isolation by a nano-filtration-based microfluidic device. Neuroscience µTongue: A Microfluidics-Based Functional Imaging Platform for the Tongue In Vivo Jisoo Han1,2, Pyonggang Choi3, Myunghwan Choi3 1Department of Biomedical Engineering, Sungkyunkwan University, 2Center for Neuroscience Imaging Research, Institute for Basic Science, 3School of Biological Sciences, Seoul National University The article introduces the µTongue (microfluidics-on-a-tongue) device for functional taste cell imaging in vivo by integrating microfluidics into an intravital imaging window on the tongue. Biology Mucociliary Epithelial Organoids from Xenopus Embryonic Cells: Generation, Culture and High-Resolution Live Imaging Hae Jin Kang1, Hye Young Kim1 1Center for Vascular Research, Institute for Basic Science We describe a simple protocol to develop mucociliary epithelial organoids from deep ectoderm cells isolated from Xenopus laevis embryos. The multipotent progenitors regenerate epithelial goblet cell precursors and allow live tracking of the initiation and progression of the cell transitions on the surface of organoids. Neuroscience Spectral Reflectometric Microscopy on Myelinated Axons In Situ Junhwan Kwon1,2, Myunghwan Choi1,2 1Department of Biomedical Engineering, Sungkyunkwan University, 2Center for Neuroscience Imaging Research, Institute for Basic Science (IBS) Here, we present a step-by-step protocol for imaging myelinated axons in a fixed brain slice using a label-free nanoscale imaging technique based on spectral reflectometry. Chemistry Liquid-cell Transmission Electron Microscopy for Tracking Self-assembly of Nanoparticles Byung Hyo Kim1,2, Junyoung Heo1,2, Won Chul Lee3, Jungwon Park1,2 1Center for Nanoparticle Research, Institute for Basic Science (IBS), 2School of Chemical and Biological Engineering, Institute of Chemical Processes, Seoul National University, 3Department of Mechanical Engineering, Hanyang University Here we introduce experimental protocols for the real-time observation of a self-assembly process using liquid-cell transmission electron microscopy. Bioengineering Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood Yang-Seok Park1, Vijaya Sunkara1, Yubin Kim1, Won Seok Lee1,2, Ja-Ryoung Han1,3, Yoon-Kyoung Cho1,4 1Department of Biomedical Engineering, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), UNIST-gil 50, Ulsan, Republic of Korea, 2Agency for Defense Development (ADD), Daejeon, Republic of Korea, 3KOGAS (Korea Gas Corporation) Research Institute, 4Center for Soft and Living Matter, Institute for Basic Science (IBS) This protocol demonstrates how to achieve femto molar detection sensitivity of proteins in 10 µL of whole blood within 30 min. This can be achieved by using electrospun nanofibrous mats integrated in a lab-on-a-disc, which offers high surface area as well as effective mixing and washing for enhanced signal-to-noise ratio.
