A high-throughput strategy for protein profiling in cell microarrays using automated image analysis
2007 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 7, no 13, 2142-2150 p.Article in journal (Refereed) Published
Advances in antibody production render a growing supply of affinity reagents for immunohistochemistry (IHC), and tissue microarray (TMA) technologies facilitate simultaneous analysis of protein expression in a multitude of tissues. However, collecting validated IHC data remains a bottleneck problem, as the standard method is manual microscopical analysis. Here we present a high-throughput strategy combining IHC on a recently developed cell microarray with a novel, automated image-analysis application (TMAx). The software was evaluated on 200 digital images of IHC-stained cell spots, by comparing TMAx annotation with manual annotation performed by seven human experts. A high concordance between automated and manual annotation of staining intensity and fraction of IHC-positive cells was found. in a limited study, we also investigated the possibility to assess the correlation between mRNA and protein levels, by using TMAx output results for relative protein quantification and quantitative real-time PCR for the quantification of corresponding transcript levels. In conclusion, automated analysis of immunohistochemically stained in vitro-cultured cells in a microarray format can be used for high-throughput protein profiling, and extraction of RNA from the same cell lines provides a basis for comparing transcription and protein expression on a global scale.
Place, publisher, year, edition, pages
2007. Vol. 7, no 13, 2142-2150 p.
antibody proteornics, automated image analysis, cell line, immunohistochemistry, tissue microarray
IdentifiersURN: urn:nbn:se:kth:diva-14253DOI: 10.1002/pmic.200700199ISI: 000248013700002ScopusID: 2-s2.0-34447566899OAI: oai:DiVA.org:kth-14253DiVA: diva2:331923
QC 201007282010-07-282010-07-282010-08-23Bibliographically approved