A gel electrophoresis loading system to prevent laboratory contamination by amplification products.

Pipet tip loading of polymerase chain reaction (PCR) and other amplification products into an electrophoresis gel represents a potential source of laboratory contamination. We have developed a prototype of the gel contamination control system (GelCCS) that enables gel loading by bottom puncture of PCR tubes. Puncture occurs within a sealed gel casing, preventing contamination of the surrounding environment. The system was designed for inexpensive manufacture so that after the results are visualized, the gel casing and PCR tubes are discarded intact with the amplification products sealed inside. We demonstrate that gel loading is reliable and that the resulting bands are equivalent in appearance to manually loaded gels.

A mill based instrument and software system for dissecting slide-mounted tissue that provides digital guidance and documentation.

BACKGROUND: Dissection of specific Areas Of Interest (AOIs) of slide-mounted tumor samples is often used to enrich for cancer cells in order to generate better signal to noise ratios in subsequent biochemical characterization. Most clinical laboratories utilize manual dissection for practical reasons and to avoid the expense and difficulties of laser microdissection systems. Unfortunately, manual methods often lack resolution and process documentation. The goal of this project was to design a dissection system for slide-mounted tissue with better precision than manual methods that also provides digital image guidance and electronic process documentation. METHODS: An instrument that is essentially a micro tissue mill was developed. It employs a specialized disposable mill bit that simultaneously dispenses liquid, cuts tissue from the slide surface, and aspirates the liquid along with the displaced tissue fragments. A software package was also developed that is capable of transferring digitally annotated AOIs between images of serially cut tissue sections to guide dissection and generate an electronic record of the process. RESULTS: The performance of this "meso" dissection system was tested using post dissection visual examination for resolution and accuracy, fluorescence based DNA quantitation for recovery efficiency, and dissection of closely situated mouse-human tissue sections followed by PCR amplification for purity determination. The minimum resolution is a dissected circle smaller than 200 microns in diameter, edge dissection accuracy is tighter than 100 microns, recovery efficiency appears greater than 95%, and recovery purity is greater than 99% relative to a different tissue located 100 microns from the dissection boundary. The system can dissect from both paraffinized and deparaffinized FFPE tissue sections that are mounted on plain glass slides, and it is compatible with DNA, RNA, and protein isolation. CONCLUSIONS: The mesodissection system is an effective alternative to manual dissection methods and is applicable for biomarker analysis of anatomical pathology samples, where enrichment of AOIs from the tissue section is helpful, but pure cell populations are not required.