October 2014: This Month in JoVE - Visualizing Infectious Disease, Transfecting with the Gene Gun, and a Novel Bioreactor System

Summary

Here's a look at what's coming up in the October 2014 issue of JoVE.

Protocol

Cultivation of Mammalian Cells Using a Single-use Pneumatic Bioreactor System

Kristina M. Obom¹, Patrick J. Cummings¹, Janelle A. Ciafardoni¹, Yasunori Hashimura², Daniel Giroux²

¹Center for Biotechnology Education, Johns Hopkins University, ²PBS Biotech, Inc.

Using a pneumatic bioreactor, we demonstrate the assembly, operation, and performance of this single-use bioreactor system for the growth of mammalian cells.

Visualizing Non-lytic Exocytosis of Cryptococcus neoformans from Macrophages Using Digital Light Microscopy

Sabriya Stukes, Arturo Casadevall

Department of Microbiology and Immunology, Albert Einstein College of Medicine

We describe how to visualize macrophage-C. neoformans (Cn) interactions in real time, with specific emphasis on the process of non-lytic exocytosis using digital light microscopy. Using this technique individually infected macrophages can be studied to ascertain various aspects of this phenomenon.

Averaging of Viral Envelope Glycoprotein Spikes from Electron Cryotomography Reconstructions using Jsubtomo

Juha T. Huiskonen, Marie-Laure Parsy, Sai Li, David Bitto, Max Renner, Thomas A. Bowden

Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford

An approach is presented for determining structures of viral membrane glycoprotein complexes using a combination of electron cryo-tomography and sub-tomogram averaging with the computational package Jsubtomo.

Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun

Jason Arsenault^1,2, Andras Nagy¹, Jeffrey T. Henderson¹, John A. O'Brien²

¹Leslie Dan Faculty of Pharmacy, University of Toronto, ²MRC-Laboratory of Molecular Biology, Cambridge, UK

Recent improvements in organotypic brain slice preparations have permitted their exploitation for biotechnological applications. Organotypic slices maintain local structural characteristics of in vivo biology, including functional synaptic connections. Here we present a regioselective biolistic delivery method to label and genetically manipulate these slices.