Summary of the July 2013 issue of BioTechniques
The July 2013 issue of BioTechniques will feature articles describing: (i) a simple method for generating patterned co-cultures of cells, (ii) a novel approach for collecting islets of Langerhans, (iii) a method for separating intact intestinal epithelium from mesenchyme, (iv) a unique methodology for immunoblotting using streptavidin, and (v) a new platform enabling the design of targeted massively parallel sequencing experiments. The July issue will also contain a Tech News article exploring the latest in transcriptional analysis techniques focusing on tools to understand transcriptional timing, as well as our regular monthly features and columns.
In tissues, cells are arranged into specific patterns to provide necessary function. Cell patterning methods and techniques provide the ability to control cell patterning in the lab, however, these techniques often involve fabrication approaches that tend to be complicated, costly, and lower-throughput. In the July issue of BioTechniques, a team of researchers from University of Toronto detail a simple, effective approach for co-culturing cells in defined locations on a plate without the need for complicated fabrication techniques. Using differential depositing of bovine serum albumin solutions and tilting of a plate, this approach presents a promising new solution for those researchers interested in medium to high-throughput patterned co-culture cell assays.
Isolation of small sections of specific tissue types can be a challenge. Two articles in the July issue present novel approaches for tissue isolation. First, in a Benchmark article, a team of researchers from University of Washington describe an innovative approach to collect pancreatic islets using a continuous flow rate in a micropipette tip such that the tissue samples settle in discrete regions of the tip. Using this approach, the authors were able to accurately and efficiently collect islets for downstream analysis.
The second article focusing on tissue isolation presents a procedure for collection of intestinal epithelium that is free of mesenchyme. Prior approaches to isolate intact intestinal epithelium have relied on long incubations, harsh chemicals, or mechanical treatments. But in the July issues of BioTechniques, researchers from Sweden detail a clever modification that enables the isolation of intestinal epithelium without harsh chemical treatment or excessive mechanical disruption. The approach, which involves no enzymes or EDTA, first requires inversion of the sample tissue, followed by ligation of one end and attachment of the other to a syringe. By then inflating the sample repeatedly, sheets of intact intestinal epithelium could be isolated by the investigators.
Loading controls typically used for quantitative immunoblotting in mammalian cells are not appropriate for Dictyostelium discoideum making protein analysis difficult. But in the July issue of BioTechniques, researchers from the United Kingdom provide a solution to this dilemma by demonstrating the use of labeled streptavidin to detect a mitochondrial protein (MCCC1), providing a robust tool for the quantitative analysis of proteins on D. discoideum Western blots.
The ability to sequence specific sections of genomes using next-generation sequencing platforms remains an important technique for researchers. Enhancing approaches for the design of targeted baits or capture probes could further assist researchers in these efforts. In a Report in July, a team of researchers from the University of Melbourne detail their new platform, called Hi-Plex, which provides a pipeline that facilitates the design and use of primers and probes for targeted massively parallel sequencing experiments. This new set of tools should further enhance the use of next-generation sequencing for scientists.
Keywords: tissue isolation, next-generation sequencing, targeted resequencing, real-time PCR, qPCR, protein analysis, Western blots, immunoblotting, streptavidin, protein localization, cell culture, patterned cell culture, cell co-culture, transcriptomics, transcriptional analysis, single cell analysis