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Open Access Technical advance

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

Nils Adey1*, Dale Emery1, Derek Bosh1, Steven Callahan3, John Schreiner3, Yang Chen1, Ann Greig4, Katherine Geiersbach2 and Robert Parry1

Author Affiliations

1 AvanSci Bio, LLC, 1290 West 2320 South, Suite D, Salt Lake City, UT 84119-1476, USA

2 Department of Pathology, ARUP Laboratories, University of Utah, 500 Chipeta Way, Salt Lake City, UT 84108, USA

3 SCI Institute, University of Utah, 72 S. Central Campus Dr., Room 3750, Salt Lake City, UT 84112, USA

4 Department of Physiology, University of Utah, 420 Chipeta Way, Suite 1700, Salt Lake City, UT 84108, USA

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BMC Clinical Pathology 2013, 13:29  doi:10.1186/1472-6890-13-29

Published: 5 November 2013

Abstract

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.

Keywords:
Slide-mounted tissue dissection; Microdissection; Mesodissection; Macrodissection; Digital pathology; Anatomic pathology tracking; FISH