Metagenomics, the study of DNA isolated from samples of naturally occurring microbial populations, is rapidly growing. Improvements to cloning and sequencing techniques are allowing researchers to study microorganisms in environmental samples, and new knowledge of species interactions and community dynamics is emerging. The identification of microorganisms in these samples is of vital importance to interpreting the results of such studies. In the January issue of Cold Spring Harbor Protocols (http://www.cshprotocols.org/TOCs/toc1_10.dtl), Annelie Wendeberg of the Helmholtz Centre for Environmental Research (http://www.ufz.de/index.php?en=13987) presents a protocol for “Fluorescence In Situ Hybridization for the Identification of Environmental Microbes.” The methods described allow the phylogenetic identification of microorganisms in environmental samples (e.g., water and sediments) by means of fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes followed by signal amplification with catalyzed reporter deposition (CARD). As one of January’s featured articles, it is freely available on the journal’s website (http://cshprotocols.cshlp.org/cgi/content/full/2010/1/pdb.prot5366).
The enzyme-linked immunospot (ELISPOT) assay is considered by many to be the gold standard for monitoring cellular immune responses. The method is highly sensitive, quantitative, easy to use, and amenable to high-throughput screening. Until recently, the ELISPOT assay has been limited to the characterization of only one effector molecule. Since the maintenance of both IFN-? and IL-2 by pathogen-specific T cells has been linked to a more favorable clinical outcome in human immunodeficiency virus (HIV) and Leishmania infections, an ELISPOT assay able to characterize both of these effector molecules would be helpful for monitoring immune responses to certain infectious agents. Nicole Bernard and colleagues from the McGill University Health Centre (http://www.mcgill.ca/hostres/investigators/bernard/) present a protocol for Dual-Color ELISPOT Assay for the Simultaneous Detection of IL-2 and/or IFN-? Secreting T Cells in the January issue of Cold Spring Harbor Protocols (http://www.cshprotocols.org/TOCs/toc1_10.dtl). As interest in multifunctional T-cell monitoring in human diseases grows, this method is likely to be extensively used. The article is freely available on the journal’s website (http://cshprotocols.cshlp.org/cgi/content/full/2010/1/pdb.prot5369).
About Cold Spring Harbor Protocols: Cold Spring Harbor Protocols (http://www.cshprotocols.org) is a monthly peer-reviewed journal of methods used in a wide range of biology laboratories. It is structured to be highly interactive, with each protocol cross-linked to related methods, descriptive information panels, and illustrative material to maximize the total information available to investigators. Each protocol is clearly presented and designed for easy use at the bench — complete with reagents, equipment, and recipe lists. Life science researchers can access the entire collection via institutional site licenses, and can add their suggestions and comments to further refine the techniques.
About Cold Spring Harbor Laboratory Press: Cold Spring Harbor Laboratory Press is an internationally renowned publisher of books, journals, and electronic media, located on Long Island, New York. Since 1933, it has furthered the advance and spread of scientific knowledge in all areas of genetics and molecular biology, including cancer biology, plant science, bioinformatics, and neurobiology. It is a division of Cold Spring Harbor Laboratory, an innovator in life science research and the education of scientists, students, and the public. For more information, visit http://www.cshlpress.com.