February 2016 - This Month in JoVE: Photoconvertible Proteins, Gold Nanoparticles, PET Principles, and Bone Marrow Microenvironments

Protocol

A Simple Method for the Size Controlled Synthesis of Stable Oligomeric Clusters of Gold Nanoparticles under Ambient Conditions

Marlon Lawrence¹, Anze Testen¹, Tilen Koklic², Oliver Smithies¹

¹Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, ²Condensed Matter Physics Department, Laboratory of Biophysics, Jozef Stefan Institute

We describe a simple method for producing highly stable oligomeric clusters of gold nanoparticles via the reduction of chloroauric acid (HAuCl₄) with sodium thiocyanate (NaSCN). The oligoclusters have a narrow size distribution and can be produced with a wide range of sizes and surface coats.

A Basic Positron Emission Tomography System Constructed to Locate a Radioactive Source in a Bi-dimensional Space

Luis Manuel Montaño-Zetina, Omar Villalobos-Mora

Physics Department, Centro de Investigacion y de Estudios Avanzados del IPN (Cinvestav)

We present a simple but well-constructed Positron Emission Tomography (PET) system and elucidate its basic working principles. The goal of this protocol is to guide the user in constructing and testing a simple PET system.

Tracking Cells in GFP-transgenic Zebrafish Using the Photoconvertible PSmOrange System

Carlo A. Beretta^1,2,3, Nicolas Dross², Ulrike Engel², Matthias Carl¹

¹Medical Faculty Mannheim, Department of Cell and Molecular Biology, Heidelberg University, ²COS and Nikon Imaging Center at the University of Heidelberg, Heidelberg University, ³University of Heidelberg, Excellenzcluster CellNetworks

We established the photoconvertible PSmOrange system as a powerful, straight-forward and cost inexpensive tool for in vivo cell tracking in GFP transgenic backgrounds. This protocol describes its application in the zebrafish model system.

Modeling Chemotherapy Resistant Leukemia In Vitro

William L. Slone*¹, Blake S. Moses*¹, Rebecca Evans¹, Debbie Piktel¹, Karen H. Martin^1,2, William Petros¹, Michael Craig¹, Laura F. Gibson^1,3

¹Alexander B. Osborn Hematopoietic Malignancy and Transplantation Program of the Mary Babb Randolph Cancer Center, Robert C. Byrd Health Sciences Center, West Virginia University School of Medicine, ²Department of Neurobiology and Anatomy, Robert C. Byrd Health Sciences Center, West Virginia University School of Medicine, ³Department of Microbiology, Immunology and Cell Biology, Robert C. Byrd Health Sciences Center, West Virginia University School of Medicine

The current report summarizes a protocol that can be utilized to model the influence of the bone marrow microenvironment niche on leukemic cells with emphasis placed on enrichment of the most chemoresistant subpopulation.