University of Bath View Institution's Website 9 articles published in JoVE Biology A High-Resolution, Single-Grain, In Vivo Pollen Hydration Bioassay for Arabidopsis thaliana Yui-Leung Lau1, Ludi Wang1, Mutian Yang1, James Doughty1 1Department of Life Sciences, University of Bath An improved method to measure pollen hydration profiles in Arabidopsis thaliana is described here. The new method offers higher resolution, is noninvasive, and is highly reproducible. The protocol represents a new tool for a finer dissection of the processes that regulate the early stages of pollination. Neuroscience Simultaneous Cryosectioning of Multiple Rodent Brains Tabitha R.F. Green1,2,3, J. Bryce Ortiz1, Jordan L. Harrison5, Jonathan Lifshitz1,3,4, Rachel K. Rowe1,3,4 1Department of Child Health, University of Arizona College of Medicine, 2Department of Biological Sciences, University of Bath, 3BARROW Neurological Institute at Phoenix Children's Hospital, 4Phoenix Veteran Affairs Healthcare System, 5Department of Basic Medical Sciences, University of Arizona College of Medicine Here, we present a protocol to freeze and section brain tissue from multiple animals as a timesaving alternative to processing single brains. This reduces staining variability during immunohistochemistry and reduces time cryosectioning and imaging. Bioengineering Hollow Fiber Bioreactors for In Vivo-like Mammalian Tissue Culture Michael P. Storm1, Ian Sorrell2, Rebecca Shipley3, Sophie Regan2, Kim A. Luetchford1, Jean Sathish2, Steven Webb4, Marianne J. Ellis1 1Department of Chemical Engineering and Centre for Regenerative Medicine, University of Bath, 2MRC Centre for Drug Safety Science and Institute of Translational Medicine, University of Liverpool, 3Mechanical Engineering, University College London, 4Department of Applied Mathematics, Liverpool John Moores University The functional behavior of cells in culture can be improved by culturing in more in vivo-like 3-dimensional culture environments16-21. This manuscript describes the set-up and operation of a hollow fiber bioreactor system for in vivo-like mammalian tissue culture. Neuroscience Primer for Immunohistochemistry on Cryosectioned Rat Brain Tissue: Example Staining for Microglia and Neurons Megan N. Evilsizor1,2, Helen F. Ray-Jones1,3, Jonathan Lifshitz1,2,4,5, Jenna Ziebell1,2 1Department of Child Health, University of Arizona College of Medicine - Phoenix, 2 This introductory level protocol describes the reagents, equipment, and techniques required to complete immunohistochemical staining of rodent brains, using markers for microglia and neuronal elements as an example. Neuroscience Examination of Synaptic Vesicle Recycling Using FM Dyes During Evoked, Spontaneous, and Miniature Synaptic Activities Sadahiro Iwabuchi1, Yasuhiro Kakazu1, Jin-Young Koh1, Kirsty M. Goodman2, N. Charles Harata1 1Department of Molecular Physiology & Biophysics, University of Iowa Carver College of Medicine, 2Department of Biology & Biochemistry, University of Bath We describe the use of styryl FM dyes to image synaptic vesicle recycling in functional nerve terminals. This protocol can be applied not only to evoked, but also spontaneous and miniature synaptic activities. The protocol expands the variety of synaptic events that can be effectively evaluated. Immunology and Infection Invasion of Human Cells by a Bacterial Pathogen Andrew M. Edwards1, Ruth C. Massey1 1Department of Biology and Biochemistry, University of Bath A general protocol for the study of invasion of host cells by a bacterial pathogen, focusing on Staphylococcus aureus and human endothelial cells. Biology Dechorionation of Medaka Embryos and Cell Transplantation for the Generation of Chimeras Sean R. Porazinski1, Huijia Wang1, Makoto Furutani-Seiki1 1Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath Due to the hard chorion and soft embryos, manipulation of medaka embryos is more involved than in zebrafish. This video shows step-by-step procedures for how to manipulate medaka embryos, including dechorionation, mounting in agarose for imaging and cell transplantation for the production of chimeras. These procedures are essential to use medaka and zebrafish in a laboratory to take full advantage of their complementary features for the genetic dissection of vertebrate genome functions. Biology Microinjection of Medaka Embryos for use as a Model Genetic Organism Sean R. Porazinski1, Huijia Wang1, Makoto Furutani-Seiki1 1Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath Medaka and zebrafish are complementary for genetic dissection of vertebrate genome functions. This protocol highlights the key points for successful microinjection into medaka embryos, an important technique for embryological and genetic analysis using medaka and zebrafish in a laboratory. Biology Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations Iwan R. Evans1, Jennifer Zanet2, Will Wood1, Brian M. Stramer2 1Department of Biology and Biochemistry, University of Bath, 2Randall Division of Cell and Molecular Biophysics, King's College London Drosophila hemocytes disperse over the entirety of the developing embryo. This protocol demonstrates how to mount and image these migrations using embryos with fluorescently labelled hemocytes.