University of San Diego 5 articles published in JoVE Biology Design and Building of a Customizable, Single-Objective, Light-Sheet Fluorescence Microscope for the Visualization of Cytoskeleton Networks Nathan Felcher1, Daisy Achiriloaie1, Brian Lee2, Ryan McGorty3, Janet Sheung1,2,4 1W.M. Keck Science Department, Scripps College, 2W.M. Keck Science Department, Claremont McKenna College, 3Department of Physics and Biophysics, University of San Diego, 4W.M. Keck Science Department, Pitzer College This protocol describes in detail how to build a single-objective, light-sheet fluorescence microscope and its usage for visualizing cytoskeleton networks. Biology Reconstituting and Characterizing Actin-Microtubule Composites with Tunable Motor-Driven Dynamics and Mechanics Mehrzad Sasanpour1, Daisy H. Achiriloaie1,2, Gloria Lee1, Gregor Leech1, Maya Hendija1, K. Alice Lindsay3, Jennifer L. Ross3, Ryan J. McGorty1, Rae M. Robertson-Anderson1 1Department of Physics and Biophysics, University of San Diego, 2W. M. Keck Science Department, Scripps College, Pitzer College, and Claremont McKenna College, 3Department of Physics, Syracuse University This paper presents protocols for engineering and characterizing tunable three-dimensional composite networks of co-entangled actin filaments and microtubules. Composites undergo active restructuring and ballistic motion, driven by myosin II and kinesin motors, and are tuned by the relative concentrations of actin, microtubules, motor proteins, and passive crosslinkers. Bioengineering Quantifying Cytoskeleton Dynamics Using Differential Dynamic Microscopy Hannah N. Verwei1, Gloria Lee2, Gregor Leech2, Irene Istúriz Petitjean3, Gijsje H. Koenderink3, Rae M. Robertson-Anderson2, Ryan James McGorty2 1Cell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht University, 2Department of Physics and Biophysics, University of San Diego, 3Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology Differential dynamic microscopy (DDM) combines features of dynamic light scattering and microscopy. Here, the process of using DDM to characterize reconstituted cytoskeleton networks by quantifying the subdiffusive and caged dynamics of particles in vimentin networks and the ballistic motion of active myosin-driven actin-microtubule composites is presented. Engineering Building Langmuir Probes and Emissive Probes for Plasma Potential Measurements in Low Pressure, Low Temperature Plasmas Peixuan Li*1, Noah Hershkowitz*1, Gregory Severn*2 1Department of Engineering Physics, University of Wisconsin, 2Department of Physics & Biophysics, University of San Diego The main goal of this work is to make it easier for research groups unfamiliar with Langmuir probes and emissive probes to use them as plasma diagnostics, especially near plasma boundaries. We do this by demonstrating how to build the probes from readily available materials and supplies. Bioengineering Gel-seq: A Method for Simultaneous Sequencing Library Preparation of DNA and RNA Using Hydrogel Matrices Gordon D. Hoople*1, Andrew Richards*2, Yan Wu2, Albert P. Pisano3, Kun Zhang2 1Department of Engineering, University of San Diego, 2Department of Bioengineering, University of California, San Diego, 3Department of Mechanical and Aerospace Engineering, University of California, San Diego Gel-seq enables researchers to simultaneously prepare libraries for both DNA- and RNA-seq at negligible added cost starting from 100 - 1000 cells using a simple hydrogel device. This paper presents a detailed approach for the fabrication of the device as well as the biological protocol to generate paired libraries.
