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Temple University

12 ARTICLES PUBLISHED IN JoVE

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Biology

Dendra2 Photoswitching through the Mammary Imaging Window
Bojana Gligorijevic 1,2, Dmitriy Kedrin 1, Jeffrey E Segall 1, John Condeelis 1,2, Jacco van Rheenen 1,2,3
1Department of Anatomy and Structural Biology, Albert Einstein College of Medicine - Yeshiva University, 2Gruss Lipper Biophotonics Center, Albert Einstein College of Medicine - Yeshiva University, 3Hubrecht Institute-KNAW and University Medical Center Utrecht

Intravital photoswitching and tracking of Dendra2-labeled tumor cells through the Mammary Imaging Window is a technique which allows us to image the metastatic behavior of tumor cells in chosen tumor microenvironments over a timescale of days.

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Biology

Single-Molecule Imaging of Nuclear Transport
Alexander Goryaynov 1, Ashapurna Sarma 1, Jiong Ma 1, Weidong Yang 1,2
1Department of Biology, Bowling Green State University, 2Center for Photochemical Sciences, Bowling Green State University

Single molecule microscopy approch provided novel insights into nuclear transport.

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Behavior

Quantifying Learning in Young Infants: Tracking Leg Actions During a Discovery-learning Task
Barbara Sargent 1, Hendrik Reimann 2, Masayoshi Kubo 3, Linda Fetters 1
1Division of Biokinesiology & Physical Therapy at the Herman Ostrow School of Dentistry, University of Southern California, 2Department of Kinesiology, Temple University, 3Department of Physical Therapy, Niigata University of Health and Welfare

A method is described in which 3-4 month old infants learn a task by discovery and their leg movements are captured to quantify the learning process.

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Chemistry

A Protocol for Safe Lithiation Reactions Using Organolithium Reagents
Michael R. Gau 1, Michael J. Zdilla 1
1Department of Chemistry, Temple University

The safe and proper use of organolithium reagents is described.

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Neuroscience

Touchscreen Sustained Attention Task (SAT) for Rats
Debra A. Bangasser 1,2, Brittany Wicks 1, David E. Waxler 1, Samantha R. Eck 1
1Psychology Department, Temple University, 2Neuroscience Program, Temple University

Sustained attention, or continuously monitoring situations for intermittent and unpredictable events, is a critical aspect of cognition. Here we detail how to test sustained attention in rats using touchscreen operant chambers. We demonstrate comparable performance in male and female rats, making this task useful for studying attention in both sexes.

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JoVE Core

Application of High-speed Super-resolution SPEED Microscopy in Live Primary Cilium
Andrew Ruba 1, Wangxi Luo 1, Weidong Yang 1
1Department of Biology, Temple University

Recently we mapped the three-dimensional (3D) spatial locations of transport routes for various proteins translocating inside primary cilia in live cells. Here this paper details the experimental setup, the process of biological samples and the data analyses for the 3D super-resolution fluorescence imaging approach newly applied in live primary cilia.

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JoVE Core

Direct Injection of a Lentiviral Vector Highlights Multiple Motor Pathways in the Rat Spinal Cord
Kathleen M. Keefe 1, Ian P. Junker 1, Imran S. Sheikh 1, Thomas J. Campion 1, George M. Smith 1
1Shriner's Pediatric Research Center, Temple University

This protocol demonstrates injection of a retrogradely transportable viral vector into rat spinal cord tissue. The vector is taken up at the synapse and transported to the cell body of target neurons. This model is suitable for retrograde tracing of important spinal pathways or targeting cells for gene therapy applications.

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Biology

Time-Resolved Fluorescence Imaging and Analysis of Cancer Cell Invasion in the 3D Spheroid Model
Louisiane Perrin 1, Theodore Tucker 1, Bojana Gligorijevic 1,2
1Bioengineering Department, Temple University, 2Cancer Biology Program, Fox Chase Cancer Center

Presented here is a protocol for the fabrication of a spheroid imaging device. This device enables dynamic or longitudinal fluorescence imaging of cancer cell spheroids. The protocol also offers a simple image processing procedure for the analysis of cancer cell invasion.

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Biology

A Bead Movement Based Computational Framework for 3-Dimensional Analysis of Biofilm Material Heterogeneity
Bettina Buttaro 1, Gillian Queisser 2
1Sol Sherry Thrombosis Research Center, Lewis Katz School of Medicine, Temple University, 2Department of Mathematics, Temple University

We describe a method for analyzing and quantifying the movement pattern of 1 µm carboxylate beads through heterogeneous bacterial biofilms. Comparison of the movement patterns can be used to quantitate differences in material properties of biofilms.

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Neuroscience

Targeting the Corticospinal Tract in Neonatal Rats with a Double-Viral Vector using Combined Brain and Spine Surgery
Rupert D. Smit 1, Thomas J. Campion III 1, Rachel L. Stingel 1, Pushti H. Shah 1, Jie Chen 1, George M. Smith 1
1Shriners Hospitals Pediatric Research Center, Lewis Katz School of Medicine, Temple University

This protocol demonstrates a novel method for applying gene therapies to subpopulations of cells in neonatal rats at postnatal ages 5-10 days by injecting an anterograde chemogenetic modifier into the somatomotor cortex and a retrogradely transportable Cre recombinase into the cervical spinal cord.

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Bioengineering

A Microphysiological System to Study Leukocyte-Endothelial Cell Interaction during Inflammation
Qingliang Yang 1, Jordan C. Langston 2, Yuan Tang 3, Balabhaskar Prabhakarpandian 4, Laurie E. Kilpatrick 5, Mohammad F. Kiani 1,2,6
1Department of Mechanical Engineering, Temple University, 2Department of Bioengineering, Temple University, 3Department of Bioengineering, University of Toledo, 4Biomedical Technology, CFD Research Corporation, 5Center for Inflammation and Lung Research, Department of Microbiology, Immunology and Inflammation, Lewis Katz School of Medicine, Temple University, 6Department of Radiation Oncology, Lewis Katz School of Medicine, Temple University

In this protocol, a biomimetic microfluid assay, which can reproduce a physiologically relevant microvascular environment and reproduce the entire leukocyte adhesion/migration cascade, is employed to study leukocyte-endothelial cell interactions in inflammatory disease.

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Bioengineering

Direct Linear Transformation for the Measurement of In-Situ Peripheral Nerve Strain During Stretching
Virginia Orozco 1, Sriram Balasubramanian 1, Anita Singh 2
1School of Biomedical Engineering, Science and Health Systems, Drexel University, 2Bioengineering Department, Temple University

This protocol implements a stereo-imaging camera system calibrated using direct linear transformation to capture three-dimensional in-situ displacements of stretched peripheral nerves. By capturing these displacements, strain induced at varying degrees of stretch can be determined informing the stretch injury thresholds that can advance the science of stretch-dependent nerve repair.

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