NUCLEIC ACID BASED MOLECULAR BIOLOGY

By Norihiro Tomita, Yasuyoshi Mori, Hidetoshi Kanda and Tsugunori Notomi.

This protocol describes the loop-mediated isothermal amplification (LAMP) method of gene amplification. It also uses a fluorescent metal indicator system to allow simple visual detection of products.

CELL AND TISSUE CULTURE

By Celeste Nelson, Jamie Inman and Mina Bissell.

A simple micromolding method is described that enables the construction of 3-D arrays of organotypic epithelial tissue structures that approximate in vivo histology. Patterned tissue arrays can be produced in 3-4 h, that undergo morphogenesis over the following one to three days.

IMAGING

By Andrew P French, Steven Mills, Ranjan Swarup, Malcolm J Bennett & Tony P Pridmore.

By Michael Sauer, Tomasz Paciorek, Eva Benková and Jiri Friml.

French et al. describe how to perform quantitative statistical colocalization on two-color confocal images, specifically of plant cells, and include a description of the alignment of the microscope. An excellent related protocol is that from Friml’s group.

BIOCHEMISTRY AND PROTEIN ANALYSIS

By Francis K.Y. Siu, Leo T.O. Lee & Billy K.C. Chow.

By Yuliang Wu, Qiang Li and Xing-Zhen Chen.

These two protocols describe different blotting methods to investigate protein-protein and protein-DNA interactions. In both techniques, proteins are firstly separated by polyacrylamide gel electrophoresis and blotted onto a membrane. Membranes are then probed to detect target interactions using either oligonucleotide probes (southwestern) or protein probes (far western).

SPECTROSCOPY AND STRUCTURAL ANALYSIS

By Wiseman, Ifa, Venter & Cooks.

DESI does not require sample preparation besides production of microtome tissue slices and does not involve ionization matrices. It allows analysis of ordinary objects or pre-processed samples under ambient conditions. For instance, it can identify and record spatial distributions of lipids and drug molecules in tissue sections.