Summary
Protocol
Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster
Saskia E.J. de Vries, Tom Clandinin
Department of Neurobiology, Stanford University
Genetically encoded optogenetic tools enable noninvasive manipulation of specific neurons in the Drosophila brain. Such tools can identify neurons whose activation is sufficient to elicit or suppress particular behaviors. Here we present a method for activating Channelrhodopsin2 that is expressed in targeted neurons in freely walking flies.
Optogenetic Activation of Zebrafish Somatosensory Neurons using ChEF-tdTomato
Ana Marie S. Palanca, Alvaro Sagasti
Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles
Optogenetic techniques have made it possible to study the contribution of specific neurons to behavior. We describe a method in larval zebrafish for activating single somatosensory neurons expressing a channelrhodopsin variant (ChEF) with a diode-pumped solid state (DPSS) laser and recording the elicited behaviors with a high-speed video camera.
Determining 3D Flow Fields via Multi-camera Light Field Imaging
Tadd T. Truscott1, Jesse Belden2, Joseph R. Nielson1, David J. Daily1, Scott L. Thomson1
1Department of Mechanical Engineering, Brigham Young University, 2Naval Undersea Warfare Center, Newport, RI
A technique for performing quantitative three-dimensional (3D) imaging for a range of fluid flows is presented. Using concepts from the area of Light Field Imaging, we reconstruct 3D volumes from arrays of images. Our 3D results span a broad range including velocity fields and multi-phase bubble size distributions.
Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging
Esperanza Mata-Martínez1, Omar José1, Paulina Torres-Rodríguez1, Alejandra Solís-López1, Ana A. Sánchez-Tusie1, Yoloxochitl Sánchez-Guevara1, Marcela B. Treviño2, Claudia L. Treviño1
1Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México, 2Math and Sciences Department, Edison State College
Intracellular Ca2+ dynamics are very important in sperm physiology and Ca2+-sensitive fluorescent dyes constitute a versatile tool to study them. Population experiments (fluorometry and stopped flow fluorometry) and single cell experiments (flow cytometry and single cell imaging) are used to track spatio-temporal [Ca2+] changes in human sperm cells.
Designing a Bio-responsive Robot from DNA Origami
Eldad Ben-Ishay, Almogit Abu-Horowitz, Ido Bachelet
DNA origami is a powerful method for fabricating precise nanoscale objects by programming the self-assembly of DNA molecules. Here, we describe how DNA origami can be utilized to design a robotic robot capable of sensing biological cues and responding by shape shifting, subsequently relayed to a desired effect.
Creating Dynamic Images of Short-lived Dopamine Fluctuations with lp-ntPET: Dopamine Movies of Cigarette Smoking
Evan D. Morris1,2,3,4, Su Jin Kim1,3, Jenna M. Sullivan1,3,4, Shuo Wang3,4, Marc D. Normandin5, Cristian C. Constantinescu6, Kelly P. Cosgrove1,2,3
1Diagnostic Radiology, Yale University, 2Psychiatry, Yale University, 3Yale PET Center, Yale University, 4Biomedical Engineering, Yale University, 5Nuclear Medicine, Massachusetts General Hospital, 6Radiological Sciences, University of California, Irvine
We present a novel PET imaging approach for capturing dopamine fluctuations induced by cigarette smoking. Subjects smoke in the PET scanner. Dynamic PET images are modeled voxel-by-voxel in time by lp-ntPET, which includes a time-varying dopamine term. The results are 'movies' of dopamine fluctuations in the striatum during smoking.
Visualization of Craniofacial Development in the sox10: kaede Transgenic Zebrafish Line Using Time-lapse Confocal Microscopy
Lisa Gfrerer, Max Dougherty, Eric C. Liao
Center for Regenerative Medicine, Massachusetts General Hospital
Visualization of experimental data has become a key element in presenting results to the scientific community. Generation of live time-lapse recording of growing embryos contributes to better presentation and understanding of complex developmental processes. This protocol is a step-by-step guide to cell labeling via photoconversion of kaede protein in zebrafish.
Disclosures
No conflicts of interest declared.