October 2012: This Month in JoVE

Summary

Here are some highlights from the October 2012 Issue of Journal of Visualized Experiments (JoVE).

Protocol

A Chitosan Based, Laser Activated Thin Film Surgical Adhesive, 'SurgiLux': Preparation and Demonstration.

L. John R. Foster, Elizabeth Karsten
Bio/Polymer Research Group, University of New South Wales

The fabrication of a novel, flexible thin film surgical adhesive from FDA approved ingredients, chitosan and indocyanine green is described. Bonding of this adhesive to collagenous tissue through a simple activation process with a low-powered infra-red laser is demonstrated.

Fabrication and Application of Rose Bengal-chitosan Films in Laser Tissue Repair

Antonio Lauto¹, Marcus Stoodley², Matthew Barton¹, John W. Morley¹, David A. Mahns¹, Leonardo Longo³, Damia Mawad^1
1Bioelectronics and Neuroscience (BENS) research group, University of Western Sydney, NSW Australia, ²Australian School of Advanced Medicine, Macquarie University, NSW Australia, ³School of Medicine, University of Siena, Italy

Sutures are usually needed to repair tissue during surgical procedures. However, their application can be problematic as they are invasive and may damage tissue. The fabrication and application methods of a novel tissue adhesive are here reported. This adhesive film is laser-activated and does not require the use of sutures.

A Galvanotaxis Assay for Analysis of Neural Precursor Cell Migration Kinetics in an Externally Applied Direct Current Electric Field

Robart Babona-Pilipos¹, Milos R. Popovic², Cindi M. Morshead^3
1Institute for Biomaterials and Biomedical Engineering, University of Toronto, ²Lyndhurst Centre, Toronto Rehabilitation Institute, ³Department of Surgery, University of Toronto

In this protocol we demonstrate how to construct custom chambers that permit the application of a direct current electric field to enable time-lapse imaging of adult brain derived neural precursor cell translocation during galvanotaxis.

Cell specific analysis of Arabidopsis leaves using fluorescence activated cell sorting

Jesper T. Grønlund¹, Alison Eyres¹, Sanjeev Kumar¹, Vicky Buchanan-Wollaston^{1, 2}, Miriam L. Gifford^{1, 2
1}School of Life Sciences, University of Warwick , ²Warwick Systems Biology, University of Warwick

A method for producing Arabidopsis leaf protoplasts that are compatible with fluorescence activated cell sorting (FACS), allowing for studies of specific cell populations. This method is compatible with any Arabidopsis line that expresses GFP in a subset of cells.

Visualization of Bacterial Toxin Induced Responses Using Live Cell Fluorescence Microscopy

Peter A. Keyel¹, Michelle E. Heid¹, Simon C. Watkins², Russell D. Salter^1
1Department of Immunology, University of Pittsburgh School of Medicine, ²Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine

Methods for purifying the cholesterol binding toxin streptolysin O from recombinant E. coli and visualization of toxin binding to live eukaryotic cells are described. Localized delivery of toxin induces rapid and complex changes in targeted cells revealing novel aspects of toxin biology.

High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease

Bryan Fiema^*, Andrew C. Harris^*, Aurelie Gomez, Praechompoo Pongtornpipat, Kelly Lamiman, Mark T. Vander Lugt, Sophie Paczesny
Pediatric Blood and Marrow Transplant Program, University of Michigan
^* These authors contributed equally

High throughput validation of multiple candidate biomarkers can be performed by sequential ELISA in order to minimize freeze/thaw cycles and use of precious plasma samples. Here, we demonstrate how to sequentially perform ELISAs for six different validated plasma biomarkers^1-3 of graft-versus-host disease (GVHD)⁴ on the same plasma sample.

Imaging Analysis of Neuron to Glia Interaction in Microfluidic Culture Platform (MCP)-Based Neuronal Axon and Glia Co-culture System

Haruki Higashimori¹, Yongjie Yang^{1, 2
1}Department of Neuroscience, Tufts University, ²Neuroscience Program, Tufts Sackler School of Graduate Biomedical Sciences

This study describes the procedures of setting up a novel neuronal axon and (astro)glia co-culture platform. In this co-culture system, manipulation of direct interaction between a single axon (and single glial cell) becomes feasible, allowing mechanistic analysis of the mutual neuron to glial signaling.

Angle-resolved Photoemission Spectroscopy At Ultra-low Temperatures

Sergey V. Borisenko¹, Volodymyr B. Zabolotnyy¹, Alexander A. Kordyuk^{1, 2}, Danil V. Evtushinsky¹, Timur K. Kim^{1, 3}, Emanuela Carleschi⁴, Bryan P. Doyle⁴, Rosalba Fittipaldi⁵, Mario Cuoco⁵, Antonio Vecchione⁵, Helmut Berger^6
1Institute for Solid State Research, IFW-Dresden, ²Institute of Metal Physics of National Academy of Sciences of Ukraine, ³Diamond Light Source LTD, ⁴Department of Physics, University of Johannesburg, ⁵CNR-SPIN, and Dipartimento di Fisica "E. R. Caianiello", Università di Salerno, ⁶Institute of Physics of Complex Matter, École Polytechnique Fédérale de Lausanne

The overall goal of this method is to determine the low-energy electronic structure of solids at ultra-low temperatures using Angle-Resolved Photoemission Spectroscopy with synchrotron radiation.

Fabrication of a Microfluidic Device for the Compartmentalization of Neuron Soma and Axons

Joseph Harris¹, Hyuna Lee¹, Behrad Vahidi¹, Christina Tu², David Cribbs³, Noo Li Jeon¹, Carl Cotman^3
1Department of Biomedical Engineering, University of California, Irvine, ²Stem Cell Research Center, University of California, Irvine, ³Institute for Brain Aging and Dementia, University of California, Irvine

In this video we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to farbricate a microfluidic device for culturing neurons.

Preparing E18 Cortical Rat Neurons for Compartmentalization in a Microfluidic Device

Joseph Harris¹, Hyuna Lee¹, Christina Tu Tu², David Cribbs³, Carl Cotman³, Noo Li Jeon^1
1Department of Biomedical Engineering, University of California, Irvine, ²Stem Cell Research Center, University of California, Irvine, ³Institute for Brain Aging and Dementia, University of California, Irvine

In this video we demonstrate the preparation of E18 Cortical Rat Neurons.

Non-plasma Bonding of PDMS for Inexpensive Fabrication of Microfluidic Devices

Joseph Harris¹, Hyuna Lee¹, Behrad Vahidi¹, Cristina Tu², David Cribbs³, Carl Cotman³, Noo Li Jeon^1
1Department of Biomedical Engineering, University of California, Irvine, ²Stem Cell Research Center, University of California, Irvine, ³Institute for Brain Aging and Dementia, University of California, Irvine

In this video we demonstrate how to use the neuron microfluidic device without plasma bonding.