Tata Institute of Fundamental Research View Institution's Website 5 articles published in JoVE Biology Reconstitution of Membrane-Tethered Minimal Actin Cortices on Supported Lipid Bilayers Darius Vasco Köster1, Abrar Bhat2, Sankarshan Talluri2, Satyajit Mayor2 1Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, 2National Centre for Biological Sciences, Tata Institute of Fundamental Research This protocol describes the formation of supported lipid bilayers and the addition of cytoskeletal filaments and motor proteins to study the dynamics of reconstituted, membrane-tethered cytoskeletal networks using fluorescence microscopy. Bioengineering A Simple Microfluidic Chip for Long-Term Growth and Imaging of Caenorhabditis elegans Jyoti Dubey1,2,3,4, Sudip Mondal1,5, Sandhya P. Koushika2 1National Centre for Biological Sciences, TIFR, 2Department of Biological Sciences, TIFR, 3InSTEM, 4Manipal Academy of Higher Education, 5Department of Mechanical Engineering, The University of Texas at Austin The protocol describes a simple microfluidic chip design and microfabrication methodology used to grow C. elegans in presence of a continuous food supply for up to 36 h. The growth and imaging device also enables intermittent long-term high-resolution imaging of cellular and sub-cellular processes during development for several days. Biochemistry RNA Blot Analysis for the Detection and Quantification of Plant MicroRNAs Varsha Tirumalai1,2, Melvin Prasad1, P. V. Shivaprasad1 1National Centre for Biological Sciences, Tata Institute of Fundamental Research, 2SASTRA University This method demonstrates use of the northern hybridization technique to detect miRNAs from total RNA extract. Developmental Biology Functional Manipulation of Maternal Gene Products Using In Vitro Oocyte Maturation in Zebrafish Elaine L. Welch*1, Celeste C. Eno*1, Sreelaja Nair2, Robin E. Lindeman3, Francisco Pelegri1 1Laboratory of Genetics, University of Wisconsin-Madison, 2Department of Biological Sciences, Tata Institute of Fundamental Research, 3Department of Genetics, Cell Biology, and Development, University of Minnesota An optimized protocol for the in vitro maturation of zebrafish oocytes used for the manipulation of maternal gene products is presented here. Bioengineering Simple Microfluidic Devices for in vivo Imaging of C. elegans, Drosophila and Zebrafish Sudip Mondal1, Shikha Ahlawat1, Sandhya P. Koushika1,2 1Neurobiology, NCBS-TIFR, 2Department of Biological Sciences, TIFR A simple microfluidic device has been developed to perform anesthetic free in vivo imaging of C. elegans, intact Drosophila larvae and zebrafish larvae. The device utilizes a deformable PDMS membrane to immobilize these model organisms in order to perform time lapse imaging of numerous processes such as heart beat, cell division and sub-cellular neuronal transport. We demonstrate the use of this device and show examples of different types of data collected from different model systems.