DUKE-NUS Graduate Medical School

5 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Visualizing Dengue Virus through Alexa Fluor Labeling

Summer Zhang¹, Hwee Cheng Tan², Eng Eong Ooi³

¹Defence Medical and Environmental Research Institute, DSO National Laboratories, ²Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School, ³Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School

Taking advantage of the advancements in fluorophore development and imaging technology, a simple method of Alexa Fluor labeling of dengue virus was devised to visualize the early interactions between virus and cell.

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.

Engineering

In Vitro Culture of Epicardial Cells From Mouse Embryonic Heart

Sindhu Ramesh^*1, Anamika Singh^*1, Dasan M. Cibi¹, Derek J. Hausenloy^1,2,3, Manvendra K. Singh^1,2

¹Program in Cardiovascular and Metabolic Disorders, Duke-NUS Graduate Medical School, ²National Heart Research Institute Singapore, National Heart Centre Singapore, ³The Hatter Cardiovascular Insititute, University College London

The epicardium is an essential source of multipotent cardiovascular progenitor cells and paracrine factors that are required for cardiovascular development and regeneration. We describe here a method to culture mouse embryonic epicardial cells.

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. Barathi^2,4,5

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.

Immunology and Infection

Use of Single Chain MHC Technology to Investigate Co-agonism in Human CD8+ T Cell Activation

Xiang Zhao¹, Maryam Hamidinia¹, Joanna Ai Ling Choo¹, Chien Tei Too^1,2, Zi Zong Ho³, Ee Chee Ren⁴, Antonio Bertoletti³, Paul A. MacAry^1,2,5, Keith G. Gould⁶, Joanna Brzostek¹, Nicholas R.J. Gascoigne^1,2,5

¹Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, ²Immunology Programme, Life Sciences Institute, National University of Singapore, ³Emerging Infectious Diseases Program, Duke-NUS Graduate Medical School, ⁴Singapore Immunology Network, A*STAR, ⁵NUS Graduate School for Integrative Sciences and Engineering (NGS), National University of Singapore, ⁶Department of Immunology, Wright-Fleming Institute, Imperial College London

This protocol describes the use of single chain MHC class I complexes to investigate molecular interactions in human CD8+ T cell activation: generation of engineered antigen presenting cells expressing single chain constructs, culture of human CD8+ T cell clone and T cell activation experiments.