A novel cell array technique for high-throughput,cell-based analysis |
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Authors: | A?Waterworth A?Hanby Email author" target="_blank">V?SpeirsEmail author |
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Institution: | (1) Molecular Medicine Unit, University of Leeds, Leeds, U.K.;(2) Academic Unit of Pathology, University of Leeds, Leeds, U.K.;(3) Molecular Medicine Unit, St. James's University Hospital, Clinical Sciences Building, LS9 7TF Leeds, U.K. |
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Abstract: | Summary Microarray technology has burgeoned over the past few years from nucleic acid-based arrays to tissue microarrays (TMAs). This
study aimed to develop a technique to incorporate cell lines into an array and to demonstrate the usefulness of this technique
by performing immunohistochemistry for β-catenin. Cell suspensions were prepared from 23 tumor cell lines. These were fixed
in formalin, suspended in agar, and embedded in paraffin to produce a cell block. A “tissue microarrayer” was used to remove
triplicate, 0.6 mm-cores from each cell block and to transfer these into a recipient paraffin block at precise coordinates.
Immunohistochemistry was used to identify cell lines positive for β-catenin. Cultured cells were successfully incorporated
into the microarray, with preservation of cell architecture and even distribution of cells within each core. A total of 18
of 69 cores (26%) were lost in processing. A total of 16 of 23 cell lines were identified as positive for membrane and cytoplasmic
β-catenin, and 6 of 23 were negative. Only one cell line was unscorable because of complete core loss. We have developed a
“cell microarray” technique for analyzing antigen expression by immunohistochemistry in multiple cell lines in a single expriment.
This novel application of microarrays permits high-throughput, cost-efficient analysis, with the potential to rapidly identify
markers with potential diagnostic and therapeutic implications in human disease. |
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Keywords: | cell lines microarray immunohistochemistry |
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