1Department of Biology, Miami University
RNA interference (RNAi)-based gene knockdown techniques are at the core of Tribolium research. Here, we provide an overview of our larval RNAi technique in Tribolium castaneum. Larval RNAi is a simple, but powerful technique that provides quick access to loss-of-function phenotypes, allowing researchers to study gene functions in diverse contexts.
Published October 13, 2014. Keywords: Molecular Biology, RNA interference, RNAi, gene knockdown, red flour beetle, Tribolium castaneum, injection, double-stranded RNA, functional analysis, teaching laboratories
1Department of Biochemistry, University of Oxford
The Drosophila egg chamber is an excellent model for studying the mechanisms of mRNA localization. In order to capture the dynamic events that underpin the processes of localization, rapid high resolution imaging of live tissue is required. Here, we present a protocol for dissection and imaging of live samples with minimal disruption.
Published February 27, 2012. Keywords: Molecular Biology, Drosophila oocytes, wide-field microscopy, cytoskeleton, RNA injection, mRNA localization
1Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah
Dying cells are extruded from epithelial tissues by concerted contraction of neighboring cells without disrupting barrier function. The optical clarity of developing zebrafish provides an excellent system to visualize extrusion in living epithelia. Here we describe methods to induce and image extrusion in the larval zebrafish epidermis at cellular resolution.
Published June 27, 2011. Keywords: Developmental Biology, Actin, Extrusion, Epithelia, Homeostasis, Zebrafish, Time-Lapse Imaging
1Department of Cell Biology, Harvard Medical School
Culturing neural explants from dissected Xenopus laevis embryos that express fluorescent fusion proteins allows for imaging of growth cone cytoskeletal dynamics.
Published October 15, 2012. Keywords: Neuroscience, Cellular Biology, Anatomy, Physiology, Growth cone, neural explant, Xenopus laevis, live cell imaging, cytoskeletal dynamics, cell culture
1Division of Cardiovascular Diseases, Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine
A rapid way to conduct immunostaining of zebrafish embryonic heart is described. Compared to the whole mount immunostaining approach, this method dramatically increases the penetration of the antibodies, which allows obtaining high resolution images that reveal cellular/subcellular structures in the heart within a much reduced processing time.
Published January 10, 2012. Keywords: Developmental Biology, Zebrafish, Danio rerio, Embryonic Heart, Cardiology, Dissection, Immunostaining