Search PubMed⌕ Search

PubMed · 12966355

Simple, inexpensive method for automating tissue microarray production provides enhanced microarray reproducibility.

Abstract

Tissue microarrays are a novel technology with the potential to impact cancer research by reducing the time, materials, and costs related to specimen-based marker validation. The process uses small cores of specimen tissue for molecular studies, maximizing the quantity of specimens that can be analyzed on a single slide and the results that can be obtained from a single antibody study. However, this process can be tedious and requires a significant time commitment for array production, particularly for the hand-produced tissue array blocks. In addition, this process has significant repetitive motions, risking repetitive stress injury for technical personnel. For these reasons, we have sought a simple, inexpensive system for automation of the existing microarray technologies. Using this system, slides containing as many as 400 specimens can be constructed in a simple and reproducible manner. Automation of the tissue microarray apparatus is accomplished by attaching two stepper motors to the micrometers of the apparatus that control array movement, and it has the advantages of standardizing the spacing between each specimen and eliminating repetitive motions by the user. A computer program is used to run the motors, allowing the user to input commands based on the desired moving distance. After assimilation of the motors, motor control boards, and corresponding program, the final product was tested and demonstrated to provide consistent, reproducible operation. Tissue microarrays were generated with specimen tissue diameters of 1.5 mm, 1.0 mm, and 0.6 mm with core densities upwards of 300 samples per slide.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Brian E Matysiak, Tracy Brodzeller, Sarah Buck, Anthony French, Cristin Counts, Brian Boorsma, Milton W Datta, Andre A Kajdacsy-Balla. 2003. Simple, inexpensive method for automating tissue microarray production provides enhanced microarray reproducibility.. https://doi.org/10.1097/00129039-200309000-00011

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

Repacking the Core of T4 lysozyme by automated design.

Automated protein redesign, as implemented in the program ORBIT, was used to redesign the core of phage T4 lysozyme. A total of 26 buried or partially buried sites in the C-terminal domain were allowed to vary both their sequence and side-chain conformation while the backbone and non-selected side-chains remained fixed. A variant with seven substitutions ("Core-7") was identified as having the most favorable energy. The redesign experiment was repeated with a penalty for the presence of methionine residues. In this case the redesigned protein ("Core-10") had ten amino acid changes. The two designed proteins, as well as the constituent single mutants, and several single-site revertants were over-expressed in Escherichia coli, purified, and subjected to crystallographic and thermal analyses. The thermodynamic and structural data show that some repacking was achieved although neither redesigned protein was more stable than the wild-type protein. The use of the methionine penalty was shown to be effective. Several of the side-chain rotamers in the predicted structure of Core-10 differ from those observed. Rather than changing to new rotamers predicted by the design process, side-chains tend to maintain conformations similar to those seen in the native molecule. In contrast, parts of the backbone change by up to 2.8A relative to both the designed structure and wild-type. Water molecules that are present within the lysozyme molecule were removed during the design process. In the redesigned protein the resultant cavities were, to some degree, re-occupied by side-chain atoms. In the observed structure, however, water molecules were still bound at or near their original sites. This suggests that it may be preferable to leave such water molecules in place during the design procedure. The results emphasize the specificity of the packing that occurs within the core of a typical protein. While point substitutions within the core are tolerated they almost always result in a loss of stability. Likewise, combinations of substitutions may also be tolerated but usually destabilize the protein. Experience with T4 lysozyme suggests that a general core repacking methodology with retention or enhancement of stability may be difficult to achieve without provision for shifts in the backbone.

Automation↗

Development, principles, and applications of automated ice fabric analyzers.

We review the recent development of automated techniques to determine the fabric and texture of polycrystalline ice. The motivation for the study of ice fabric is first outlined. After a brief introduction to the relevant optical concepts, the classic manual technique for fabric measurement is described, along with early attempts at partial automation. Then, the general principles behind fully automated techniques are discussed. We describe in some detail the similarities and differences of the three modern instruments recently developed for ice fabric studies. Next, we discuss briefly X-ray, radar, and acoustic techniques for ice fabric characterization. We also discuss the principles behind automated optical techniques to measure fabric in quartz rock samples. Finally, examples of new applications that have been facilitated by the development of the ice fabric instruments are presented.

Automation↗

Automated large-volume sample stacking procedure to detect labeled peptides at picomolar concentration using capillary electrophoresis and laser-induced fluorescence detection.

We have developed an automated large-volume sample stacking (LVSS) procedure to detect fluorescein isothiocyanate-labeled peptides in the picomolar range. The injection duration is 10 min at 50 mbar to fill 62% of the capillary volume to the detection cell. The calculated limit of detection (S/N=3), filling 1% of the capillary volume, is 74 pM for bradykinin and 45 pM for L-enkephalin with samples diluted in water and analyzed in a 50 mM borate buffer, pH 9.2. With the automated LVSS system, the limits of detection are 7 pM for bradykinin, 3 pM for L-enkephalin and 2 pM for substance P. LVSS is shown to be quantitative from 500 to 10 pM.

Automation↗