Volume 10 Issue 8, August 2015

This protocol generates oligodendrocyte progenitor cells from human pluripotent stem cells. The cells can subsequently be isolated by cell sorting for myelination studies, or they can be terminally differentiated to MBP⁺ oligodendrocytes.

This protocol describes how to use the chicken embryo to assess virulence of the human bacterial pathogen Listeria monocytogenes, but it could also be extended to other microbial pathogens.

This protocol generates and characterizes dispersed cells from Pseudomonas aeruginosa biofilms in vitro and in vivo by reducing the intracellular c-di-GMP content. Dispersed biofilm cells have an important role in transmission of infections.

This protocol combines the use of a custom-fabricated microfluidic device and time-lapse microscopy with automated image analysis to measure cell signaling in single yeast cells on a high-throughput scale.

Roberts et al. describe how to measure transcriptional activity using nuclear run-on RT-qPCR assays. The approach is ideal for analyzing a single gene (or small number of genes) or for follow-up or validation of genome-wide analyses.

This protocol is for high-throughput sequencing–RNA affinity profiling (HiTS-RAP). The technique uses an unmodified Illumina GAIIx sequencer to analyze millions of RNA-protein binding affinities in a single run.

This protocol describes how to prepare plants for light-sheet microscopy. Seeds or seedlings are suspended in low-gelling-temperature agarose or they grow in Phytagel within glass capillaries or FEP tubes. This allows long-term growth and imaging of tissues with the Lightsheet Z.1 microscope.

This protocol describes a detailed method for superresolution imaging of plant tissues by structured illumination microscopy (SIM). Details include microscope calibration, tissue preparation, image acquisition and evaluation of SIM images.

This protocol describes a syngeneic orthotopic hepatocellular carcinoma (HCC) model in immunocompetent mice with liver cirrhosis induced by carbon tetrachloride. This model recapitulates the key features of the necroinflammatory, fibrotic and angiogenic microenvironment of human HCC.