A novel cell array technique for high-throughput, cell-based analysis.
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 beta-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 beta-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 beta-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 experiment. 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.
Waterworth
A
A
Molecular Medicine Unit and Academic Unit of Pathology, University of Leeds, Leeds, U.K.
Hanby
A
A
Speirs
V
V
D014408:Biomarkers, Tumor / Q000032:analysis; D045744:Cell Line, Tumor / Q000737:chemistry* / Q000166:cytology; D006652:Histological Techniques; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D025202:Molecular Diagnostic Techniques; D051176:beta Catenin / Q000032:analysis*
D014408:Biomarkers, Tumor; D051176:beta Catenin