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4 ARTICLES PUBLISHED IN JoVE

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Biology

Isolation and Culture of Primary Mouse Keratinocytes from Neonatal and Adult Mouse Skin
Fengwu Li 1, Christopher A. Adase 1, Ling-juan Zhang 1
1Department of Dermatology, School of Medicine, UC San Diego

Epidermal keratinocytes form a functional skin barrier and are positioned at the front line of host defense against external environmental insults. Here we describe methods for isolation and primary culture of epidermal keratinocytes from neonatal and adult mouse skin, and induction of terminal differentiation and UVB-triggered inflammatory response from keratinocytes.

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Neuroscience

Generation and Long-term Maintenance of Nerve-free Hydra
Cassidy M. Tran 1, Sharon Fu 1, Trevor Rowe 2, Eva-Maria S. Collins 1,2
1Division of Biological Sciences, UC San Diego, 2Physics Department, UC San Diego

Through a double treatment with colchicine, a plant-derived toxin that kills dividing cells, nerve-free Hydra vulgaris can be generated. These Hydra cannot feed or egest on their own. This paper describes an improved method for long-term maintenance of nerve free Hydra vulgaris in the laboratory.

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Biology

Planarian Scrunching as a Quantitative Behavioral Readout for Noxious Stimuli Sensing
Ziad Sabry 1, Christina Rabeler 1, Danielle Ireland 1, Kevin Bayingana 1, Eva-Maria S. Collins 1,2
1Biology Department, Swarthmore College, 2Physics Department, UC San Diego

Freshwater planarians exhibit three gaits (gliding, peristalsis, and scrunching) that are distinguishable by quantitative behavioral analysis. We describe a method to induce scrunching using various noxious stimuli, quantification thereof, and distinction from peristalsis and gliding. Using gene knockdown, we demonstrate the specificity of scrunching as a quantitative phenotypic readout.

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Chemistry

Multimodal Nonlinear Hyperspectral Chemical Imaging Using Line-Scanning Vibrational Sum-Frequency Generation Microscopy
Jackson C. Wagner 1, Bin Yang 1,2, Zishan Wu 1, Wei Xiong 1,3,4
1Department of Chemistry and Biochemistry, UC San Diego, 2State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 3Materials Science and Engineering Program, UC San Diego, 4Department of Electrical and Computer Engineering, UC San Diego

A multimodal, rapid hyperspectral imaging framework was developed to obtain broadband vibrational sum-frequency generation (VSFG) images, along with brightfield, second harmonic generation (SHG) imaging modalities. Due to the infrared frequency being resonant with molecular vibrations, microscopic structural and mesoscopic morphology knowledge is revealed of symmetry-allowed samples.

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