Nanyang Technological University View Institution's Website 40 articles published in JoVE Biochemistry A Nanobar-Supported Lipid Bilayer System for the Study of Membrane Curvature Sensing Proteins in vitro Xinwen Miao*1, Jiawei Wu*1,2, Wenting Zhao1,3 1School of Chemistry, Chemical Engineering, and Biotechnology, Nanyang Technological University, 2State Key Laboratory Breeding Base of Green Chemistry Synthesis Technology, College of Chemical Engineering, Zhejiang University of Technology, 3Institute for Digital Molecular Analytics and Science, Nanyang Technological University Here, a nanobar-supported lipid bilayer system is developed to provide a synthetic membrane with a defined curvature that enables the characterization of proteins with curvature sensing ability in vitro. Bioengineering Spatio-Temporal In Vivo Imaging of Ocular Drug Delivery Systems using Fiberoptic Confocal Laser Microendoscopy Su Yin Chaw1,2, Tina Tzee Ling Wong3,4, Subbu Venkatraman2,5, Ann-Marie Chacko1 1Laboratory for Translational and Molecular Imaging, Cancer and Stem Cell Biology Programme, Duke-NUS Medical School, 2School of Materials Science and Engineering, Nanyang Technological University, 3Singapore National Eye Centre, 4Singapore Eye Research Institute, 5Material Science & Engineering, National University of Singapore We present a protocol for the use of fiberoptic confocal laser microendoscopy (CLM) to non-invasively study the spatio-temporal distribution of liposomes in the eye after subconjunctival injection. Bioengineering Segmenting Growth of Endothelial Cells in 6-Well Plates on an Orbital Shaker for Mechanobiological Studies Kuin T. Pang*1,2, Mean Ghim*1, Mehwish Arshad1, Xiaomeng Wang2,3,4, Peter D. Weinberg1 1Department of Bioengineering, Imperial College London, 2Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A*STAR), 3Lee Kong Chian School of Medicine, Nanyang Technological University Singapore, 4Singapore Eye Research Institute, The Academia This protocol describes a coating method to restrict endothelial cell growth to a specific region of a 6-well plate for shear stress application using the orbital shaker model. Environment Visualization of Flow Field Around a Vibrating Pipeline Within an Equilibrium Scour Hole Dawei Guan1,2, Yee-Meng Chiew2, Maoxing Wei3, Shih-Chun Hsieh4 1Key Laboratory of Coastal Disaster and Defense, Ministry of Education, Hohai University, 2School of Civil and Environmental Engineering, Nanyang Technological University, 3Ocean College, Zhejiang University, 4AXESEA Information Technology Co. LTD The goal of the protocol is to enable visualization of the detailed flow fields and determination of the near-boundary shear and normal stresses within an equilibrium scour hole induced by a vibrating pipeline. Bioengineering Pulsed Laser Diode-Based Desktop Photoacoustic Tomography for Monitoring Wash-In and Wash-Out of Dye in Rat Cortical Vasculature Sandeep Kumar Kalva1, Paul Kumar Upputuri1, Praveenbalaji Rajendran1, Rhonnie Austria Dienzo1, Manojit Pramanik1 1School of Chemical and Biomedical Engineering, Nanyang Technological University A compact pulsed laser diode-based desktop photoacoustic tomography (PLD-PAT) system is demonstrated for high-speed dynamic in vivo imaging of small animal cortical vasculature. Genetics Genome Editing in Mammalian Cell Lines using CRISPR-Cas Kaiwen Ivy Liu1, Norfala-Aliah Binte Sutrisnoh1, Yuanming Wang1,2, Meng How Tan1,2 1Genome Institute of Singapore, Agency for Science Technology and Research, 2School of Chemical and Biomedical Engineering, Nanyang Technological University CRISPR-Cas is a powerful technology to engineer the complex genomes of plants and animals. Here, we detail a protocol to efficiently edit the human genome using different Cas endonucleases. We highlight important considerations and design parameters to optimize editing efficiency. Engineering Optimization, Test and Diagnostics of Miniaturized Hall Thrusters Jian W. M. Lim1, Igor Levchenko1,2, Muhammad W. A. B. Rohaizat1, Shiyong Huang1, Luxiang Xu1, Yu Fei Sun1, George C. Potrivitu1, Jen S. Yee1, Roysmond Z. W. Sim1, Youmei Wang3, Svitlana Levchenko1, Kateryna Bazaka2,1, Shuyan Xu1 1Plasma Sources and Applications Centre, National Institute of Education, Nanyang Technological University, 2School of Chemistry, Physics, Mechanical Engineering, Queensland University of Technology, 3Department of Physics, School of Science, Hangzhou Dianzi University Here, we present a protocol to test and optimize space propulsion systems based on miniaturized Hall-type thrusters. Engineering Microparticle Manipulation by Standing Surface Acoustic Waves with Dual-frequency Excitations Yufeng Zhou1, Yannapol Sriphutkiat1 1School of Mechanical and Aerospace Engineering, Nanyang Technological University A protocol for manipulating the microparticles in a microfluidic channel with a dual-frequency excitation is presented. Biology Co-immunoprecipitation Assay Using Endogenous Nuclear Proteins from Cells Cultured Under Hypoxic Conditions Xiaofeng Zheng1,2, Calvin Qing Wei Ho1, Xiaowei Zheng3, Kian Leong Lee4, Katarina Gradin5, Teresa S. Pereira3, Per-Olof Berggren1,2,3, Yusuf Ali1,2 1Lee Kong Chian School of Medicine, Nanyang Technological University, 2Singapore Eye Research Institute (SERI), Singapore General Hospital, 3The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, 4Cancer and Stem Cell Biology Program, Duke-NUS Medical School, 5Department of Cell and Molecular Biology, Karolinska Institutet Here we describe a co-immunoprecipitation protocol to study protein-protein interactions between endogenous nuclear proteins under hypoxic conditions. This method is suitable for demonstration of the interactions between transcription factors and transcriptional co-regulators at hypoxia. Chemistry Synthesis of Substrate-Bound Au Nanowires Via an Active Surface Growth Mechanism Xinglong Wang*1, Xuesong Wu*1, Jiating He2, Xiaolin Tao1, Hongyan Li1, Gui Zhao1, Yawen Wang1, Hongyu Chen1,2 1Institute of Advanced Synthesis, School of Chemistry and Molecular Engineering, Jiangsu National Synergetic Innovation Center for Advanced Materials, Nanjing Tech University, 2Chemistry and Biological Chemistry, Nanyang Technological University We report a solution-based method to synthesize substrate-bound Au nanowires. By tuning the molecular ligands used during the synthesis, the Au nanowires can be grown from various substrates with different surface properties. Au nanowire-based nanostructures can also be synthesized by adjusting the reaction parameters. Chemistry Synthesis of Core-shell Lanthanide-doped Upconversion Nanocrystals for Cellular Applications Xiangzhao Ai1, Linna Lyu1, Jing Mu1, Ming Hu1, Zhimin Wang1, Bengang Xing1,2 1Division of Chemistry and Biological Chemistry, School of Physical & Mathematical Sciences, Nanyang Technological University, 2Institute of Materials Research & Engineering, Agency for Science, Technology and Research (A*STAR) A protocol is presented for the synthesis of core-shell lanthanide-doped upconversion nanocrystals (UCNs) and their cellular applications for channel protein regulation upon near-infrared (NIR) light illumination. Bioengineering Hand-held Clinical Photoacoustic Imaging System for Real-time Non-invasive Small Animal Imaging Kathyayini Sivasubramanian1, Vijitha Periyasamy1, Manojit Pramanik1 1School of Chemical and Biomedical Engineering, Nanyang Technological University A clinical handheld photoacoustic imaging system will be demonstrated for real-time non-invasive small animal imaging. Genetics Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids Desiree-Faye Kaixin Toh*1, Kiran M. Patil*1, Gang Chen1 1Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University We report the protocols for the synthesis and purification of Peptide Nucleic Acid (PNA) oligomers incorporating modified residues. The biochemical and biophysical methods for the characterization of the recognition of RNA duplexes by the modified PNAs are described. Biology Quantitative Localization of a Golgi Protein by Imaging Its Center of Fluorescence Mass Hieng Chiong Tie1, Bing Chen1, Xiuping Sun1, Li Cheng2,3, Lei Lu1 1School of Biological Sciences, Nanyang Technological University, 2Bioinformatics Institute, 3School of Computing, National University of Singapore The precise localization of Golgi residents is essential for understanding the cellular functions of the Golgi. However, conventional optical microscopy is unable to resolve the sub-Golgi structure. Here we describe the protocol for a conventional microscopy based super-resolution method to quantitatively determine the sub-Golgi localization of a protein. Immunology and Infection A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva Tanima Bose1, Aihua Hou2, Ryan Lee2, Louis Tong2,3,4,5, K. George Chandy1 1Lee Kong Chian School of Medicine, Nanyang Technological University, 2Singapore Eye Research Institute, 3Singapore National Eye Center, 4Duke-NUS Medical School, 5Yong Loo Lin School of Medicine The exposed normal ocular surface consists of cornea and conjunctiva. Epithelial cells, goblet cells and immune cells are present in the conjunctiva. Here, a non-invasive, technique of impression cytology is described using an impression cytology device and flow cytometry to analyze immune cells in the conjunctiva. Medicine Fluorescent Dye Labeling of Erythrocytes and Leukocytes for Studying the Flow Dynamics in Mouse Retinal Circulation Rupesh Agrawal*1,2,3, Praveen Kumar Balne*2, Sai Bo Bo Tun2, Yeo Sia Wey2, Neha Khandelwal1, Veluchamy A. Barathi2,4,5 1National Healthcare Group Eye Institute, Tan Tock Seng Hospital, 2Singapore Eye Research Institute (SERI), Singapore National Eye Center, 3School of Material Science and Engineering, Nanyang Technological University, 4Department of Ophthalmology, Yong Loo Lin School of Medicine, National University Health Systems, National University of Singapore, 5Ophthalmology Academic Clinical Research Program, DUKE-NUS Graduate Medical School Live-cell imaging of the labeled blood cells in ocular circulation can provide information about inflammation and ischemia in diabetic retinopathy and age-related macular degeneration. A protocol to label blood cells and image the labeled cells in the retinal circulation is described. Bioengineering Switchable Acoustic and Optical Resolution Photoacoustic Microscopy for In Vivo Small-animal Blood Vasculature Imaging Mohesh Moothanchery1, Arunima Sharma1, Manojit Pramanik1 1School of Chemical and Biomedical Engineering, Nanyang Technological University Here a switchable acoustic resolution (AR) and optical resolution (OR) photoacoustic microscopy (AR-OR-PAM) system capable of both high resolution imaging at shallow depth and low resolution deep tissue imaging on the same sample in vivo is demonstrated. Bioengineering A High-performance Compact Photoacoustic Tomography System for In Vivo Small-animal Brain Imaging Paul Kumar Upputuri1, Vijitha Periyasamy1, Sandeep Kumar Kalva1, Manojit Pramanik1 1School of Chemical and Biomedical Engineering, Nanyang Technological University A compact pulsed-laser diode-based photoacoustic tomography (PLD-PAT) system for high-speed in vivo brain imaging in small animals is demonstrated. Immunology and Infection A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue Aihua Hou1,2, Tanima Bose3, K. George Chandy3, Louis Tong1,2,4,5 1Singapore Eye Research Insitute, A Member of SingHealth, 2DUKE-National University of Singapore Medical School, 3Lee Kong Chian School of Medicine, Nanyang Technological University, 4Singapore National Eye Center, 5Yong Loo Lin School of Medicine, National University of Singapore This report describes a method to induce chronic experimental autoimmune dry eye in Lewis rats through immunization with an emulsion of rat lacrimal gland extract, ovalbumin, and complete Freund's adjuvant, followed by the injection of lacrimal gland extract and ovalbumin into the forniceal subconjunctiva and lacrimal glands six weeks later. Biochemistry LERLIC-MS/MS for In-depth Characterization and Quantification of Glutamine and Asparagine Deamidation in Shotgun Proteomics Xavier Gallart-Palau*1, Aida Serra*1, Siu Kwan Sze1 1Division of Structural Biology and Biochemistry, School of Biological Sciences, Nanyang Technological University Here we present a step-by-step protocol of the long-length electrostatic repulsion-hydrophilic interaction chromatography-tandem mass spectrometry (LERLIC-MS/MS) method. This is a novel methodology that enables for the first time quantification and characterization of the glutamine and asparagine deamidation isoforms by shotgun proteomics. Medicine Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses Chi Ching Goh1,2, Jackson LiangYao Li1, David Becker3, Wolfgang Weninger4,5,6, Veronique Angeli2,7, Lai Guan Ng1,2,5,8 1Singapore Immunology Network (SIgN), Agency for Science, Technology and Research (A*STAR), Biopolis, 2Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 3Lee Kong Chian School of Medicine, Nanyang Technological University, 4Centenary Institute for Cancer Medicine and Cell Biology, 5Discipline of Dermatology, University of Sydney, 6Department of Dermatology, Royal Prince Alfred Hospital, 7LSI Immunology Programme, National University of Singapore, 8School of Biological Sciences, Nanyang Technological University This protocol describes the induction of an ischemia-reperfusion (IR) model on mouse ear skin using magnet clamping. Using a custom-built intravital imaging model, we study in vivo inflammatory responses post-reperfusion. The rationale behind the development of this technique is to extend the understanding of how leukocytes respond to skin IR injury. Bioengineering Automated Robotic Dispensing Technique for Surface Guidance and Bioprinting of Cells Ramya Bhuthalingam1, Pei Q. Lim1, Scott A. Irvine1, Subbu S. Venkatraman1 1School of Materials Science and Engineering, Nanyang Technological University This protocol describes a bioprinting methodology using an automated robotic depositing system that incorporates etched topographical guidance cues with the precision deposition of a cell bearing hydrogel bioink. The printed cells are directly delivered to the etched features and are able to sense and orientate with them. Bioengineering Extraction of Plant-based Capsules for Microencapsulation Applications Michael G. Potroz1,2, Raghavendra C. Mundargi1,2, Jae Hyeon Park1,2, Ee-Lin Tan1,2, Nam-joon Cho1,2,3 1School of Materials Science and Engineering, Nanyang Technological University, 2Centre for Biomimetic Sensor Science, Nanyang Technological University, 3School of Chemical and Biomedical Engineering, Nanyang Technological University This protocol/manuscript describes a streamlined process for the production of sporopollenin exine capsules (SECs) from Lycopodium clavatum spores and for the loading of hydrophilic compounds into these SECs. Immunology and Infection Isolation and Activation of Murine Lymphocytes Jun Feng Lim1, Heidi Berger1, I-hsin Su1 1School of Biological Sciences, College of Science, Nanyang Technological University Lymphocytes are the major players in adaptive immune responses. Here, we present a lymphocyte purification protocol to determine the physiological functions of the desired molecules in lymphocyte activation in vitro and in vivo. The described experimental procedures are suitable for comparing functional capacities between control and genetically modified lymphocytes. Chemistry 1,3,5-Triphenylbenzene and Corannulene as Electron Receptors for Lithium Solvated Electron Solutions Kim Seng Tan1,2, Andrey V. Lunchev2, Mihaiela C. Stuparu2,3, Andrew C. Grimsdale2, Rachid Yazami1,2 1Energy Research Institute (ERI@N), Nanyang Technological University, 2School of Materials Science and Engineering, Nanyang Technological University, 3School of Physical and Mathematical Sciences, Nanyang Technological University The authors report on conductivity studies carried out on lithium solvated electron solutions (LiSES) prepared using 1,3,5-triphenylbenzene (TPB) and corannulene as electron receptors. Neuroscience Insect-machine Hybrid System: Remote Radio Control of a Freely Flying Beetle (Mercynorrhina torquata) T. Thang Vo Doan1, Hirotaka Sato1 1School of Mechanical and Aerospace Engineering, Nanyang Technological University This protocol describes the process of constructing an insect-machine hybrid system and carrying out wireless electrical stimulation of the flight muscles required to control the turning motion of a flying insect. Bioengineering Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering Hitomi Shirahama1, Supriya K. Kumar1, Won-Yong Jeon1, Myung Hee Kim1, Jae Ho Lee1, Soon Seng Ng1, Seyed R. Tabaei1, Nam-Joon Cho1,2 1School of Materials Science and Engineering, Nanyang Technological University, 2School of Chemical and Biomedical Engineering, Nanyang Technological University This manuscript presents a detailed protocol for the fabrication of an emerging three-dimensional hepatocyte culture platform, the inverted colloidal crystal scaffold, and the concomitant techniques to assess hepatocyte behavior. The size-controllable pores, interconnectivity and ability to conjugate extracellular matrix proteins to the poly(ethylene glycol) (PEG) scaffold enhance Huh-7.5 cell performance. Biology Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture Qianfan Bai*1, Zhiqiang Bai*1, Shaohai Xu3, Lei Sun1,2 1Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, 2Institute of Molecular and Cell Biology, Singapore, 3Division of Bioengineering, School of Chemical & Biomedical Engineering, Nanyang Technological University An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes. Bioengineering Microfluidic Buffer Exchange for Interference-free Micro/Nanoparticle Cell Engineering Hui Min Tay1, David C. Yeo2, Christian Wiraja2, Chenjie Xu2,3, Han Wei Hou1 1Lee Kong Chian School of Medicine, Nanyang Technological University, 2School of Chemical and Biomedical Engineering, Nanyang Technological University, 3NTU-Northwestern Institute of Nanomedicine, Nanyang Technological University This protocol describes the use of an inertial microfluidics-based buffer exchange strategy to purify micro/nanoparticle engineered cells with efficient depletion of unbound particles. Engineering Compact Lens-less Digital Holographic Microscope for MEMS Inspection and Characterization Thomas Bourgade1, Sun Jianfei1, Zhaomin Wang1, Rosmin Elsa2, Anand Asundi1,2 1School of Mechanical and Aerospace Engineering, Center of Laser and Optical Engineering, Nanyang Technological University, 2 We present a compact reflection digital holographic system (CDHM) for inspection and characterization of MEMS devices. A lens-less design using a diverging input wave providing natural geometrical magnification is demonstrated. Both static and dynamic studies are presented. Bioengineering Synthesis of Gold Nanoparticle Integrated Photo-responsive Liposomes and Measurement of Their Microbubble Cavitation upon Pulse Laser Excitation Malathi Mathiyazhakan1, Weiwei Chan2, Claus-Dieter Ohl2, Chenjie Xu1,3 1School of Chemical and Biomedical Engineering, Nanyang Technological University, 2Division of Physics, School of Physical and Mathematical Sciences, Nanyang Technological University, 3NTU-Northwestern Institute of Nanomedicine, Nanyang Technological University This protocol describes a simple preparation method for gold nanoparticle integrated photo-responsive liposomes with the commercially available materials. It also shows how to measure the microbubble cavitation process of the synthesized liposomes upon the treatment of pulsed laser. Bioengineering In Situ Mapping of the Mechanical Properties of Biofilms by Particle-tracking Microrheology Su C. Chew1,2, Scott A. Rice2,3,4,5, Staffan Kjelleberg2,3,4,6, Liang Yang2,3 1Interdisciplinary Graduate School, Nanyang Technological University, 2Singapore Centre on Environmental Life Sciences Engineering, Nanyang Technological University, 3School of Biological Sciences, Nanyang Technological University, 4Centre for Marine Bio-Innovation, University of New South Wales, 5School of Biological, Earth and Environmental Sciences, University of New South Wales, 6School of Biotechnology and Biomolecular Sciences Sciences, University of New South Wales Particle-tracking microrheology investigates the viscoelasticity of materials. Here, the technique is used to determine the viscoelasticity, creep compliance and effective crosslinking roles of different matrix components of a bacterial biofilm. The matrix consists of polymeric substances secreted by the bacteria and its components determine biofilm structure and mechanical properties. Bioengineering Biomembrane Fabrication by the Solvent-assisted Lipid Bilayer (SALB) Method Seyed R. Tabaei1,2, Joshua A. Jackman1,2, Minchul Kim1,2, Saziye Yorulmaz1,2, Setareh Vafaei1,2, Nam-Joon Cho1,2,3 1School of Materials Science and Engineering, Nanyang Technological University, 2Centre for Biomimetic Sensor Science, Nanyang Technological University, 3School of Chemical and Biomedical Engineering, Nanyang Technological University We present an experimental protocol to form a supported lipid bilayer on solid substrates without using lipid vesicles. We demonstrate a one-step method to form a lipid bilayer on silicon dioxide and gold as well as supported membranes with cholesterol-enriched domain for various biological applications. Bioengineering A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening Yong Yu1, Siu Yee New1, Jiaxian Lin2, Xiaodi Su1, Yen Nee Tan1 1Institute of Materials Research and Engineering, Agency for Science, Technology and Research (A*STAR), 2School of Chemical & Biomedical Engineering, Nanyang Technological University A protocol for small molecular drug screening based on in-situ synthesis of ultrasmall fluorescent gold nanoclusters (Au NCs) using drug-loaded protein as template is presented. This method is simple to determine the binding affinity of drugs to a target protein by a visible fluorescent signal emitted from the protein-templated Au NCs. Chemistry Using Polystyrene-block-poly(acrylic acid)-coated Metal Nanoparticles as Monomers for Their Homo- and Co-polymerization Yawen Wang1, Xiohui Song1, Hong Wang1, Hongyu Chen1 1Division of Chemistry and Biological Chemistry, Nanyang Technological University We report protocols for “polymerizing” various types of polymer-encapsulated metal nanoparticles into long chains of “homo-“ and “co-polymers”. Bioengineering Design and Construction of Artificial Extracellular Matrix (aECM) Proteins from Escherichia coli for Skin Tissue Engineering Pearlie S. J. Low1, Monica S. Tjin1, Eileen Fong1 1School of Materials Science and Engineering, Nanyang Technological University Recombinant technologies have enabled material designers to create novel artificial proteins with customized functionalities for tissue engineering applications. For example, artificial extracellular matrix proteins can be designed to incorporate structural and biological domains derived from native ECMs. Here, we describe the construction and purification of aECM proteins containing elastin-like repeats. Chemistry Sedimentation Equilibrium of a Small Oligomer-forming Membrane Protein: Effect of Histidine Protonation on Pentameric Stability Wahyu Surya1, Jaume Torres1 1School of Biological Sciences, Nanyang Technological University Sedimentation equilibrium (SE) can be used to study protein-protein interactions in a physiological environment. This manuscript describes the use of this technique to determine the effect of pH on the stability of a homo-pentamer formed by the small hydrophobic (SH) protein encoded by the human syncytial respiratory virus (hRSV). Developmental Biology An Optogenetic Approach for Assessing Formation of Neuronal Connections in a Co-culture System Colin T. E. Su*1, Su-In Yoon*2, Guillaume Marcy*1, Eunice W. M. Chin1, George J. Augustine2, Eyleen L. K. Goh1 1Neuroscience & Behavioral Disorders, Duke-NUS Graduate Medical School, 2Lee Kong Chian School of Medicine, Nanyang Technological University A protocol to generate a co-culture system consisting of neurons derived from induced pluripotent stem cells (iPSCs), primary cortical neurons and astrocytes is described. This co-culture system allows detection of the formation of synaptic contacts and circuits between new, iPSC-derived neurons and pre-existing cortical neurons expressing channelrhodopsin-2. Bioengineering Preparation of Mica Supported Lipid Bilayers for High Resolution Optical Microscopy Imaging Artur Matysik1, Rachel S. Kraut1 1School of Biological Sciences, Nanyang Technological University We present a method of preparing mica supported lipid bilayers for high resolution microscopy. Mica is transparent and flat on an atomic scale, but rarely used in imaging because of handling difficulties; our preparation results in even deposition of the mica sheet, and reduces the material used in bilayer preparation. Biology Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE) Heike Summer1, René Grämer2, Peter Dröge1 1School of Biological Sciences, Nanyang Technological University, Singapore - NTU, 2Singapore-MIT Alliance for Reserach and Technology (SMART) Denaturing urea polyacrylamide gel electrophoresis is used to separate single-stranded DNA or RNA up to a limit of 500 nucleotides. Urea in combination with heat denatures samples and unstructured single strands migrate within the gel matrix according to their molecular weight.