Abstract
The IN Cell Analyzer 1000 possesses several distinguishing features that make it a valuable tool in research today. This fully automated high content screening (HCS) system introduced quantitative fluorescent microscopy with computerized image analysis for use in cell-based analysis. Previous studies have focused on live cell assays, where it has proven to be a powerful and robust method capable of providing reproducible, quantitative data. Using HCS as a tool to investigate antigen expression in duodenal biopsies, we developed a novel approach to tissue positioning and mapping. We adapted IN Cell Analyzer 1000’s image acquisition and analysis software for the investigation of tissue transglutaminase (tTG) and smooth muscle alpha-actin (SM α-actin) staining in paraffin-embedded duodenal tissue sections from celiac patients and healthy controls. These innovations allowed a quantitative analysis of cellular structure and protein expression. The results from routine biopsy material indicated the intensity of protein expression was altered in celiac disease compared to normal biopsy material.
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References
Ghosh RN, DeBiasio R, Hudson CC, Ramer ER, Cowan CL, Oakley RH (2005) Quantitative cell-based high-content screening for vasopressin receptor agonists using transfluor technology. J Biomol Screen 10(5):476–484. doi:10.1177/1087057105274896
Ghosh RN, Lapets O, Haskins JR (2007) Characteristics and value of directed algorithms in high content screening. Methods Mol Biol (Clifton, NJ) 356:63–81
Mohamed BM, Movia D, Knyazev A, Langevin D, Davies AM, Prina-Mello A, Volkov Y (2013) Citrullination as early-stage indicator of cell response to single-walled carbon nanotubes. Sci Rep 3:1124. doi:10.1038/srep01124
Mohamed BM, Verma NK, Davies AM, McGowan A, Crosbie-Staunton K, Prina-Mello A, Volkov Y (2012) Citrullination of proteins: a common post-translational modification pathway induced by different nanoparticles in vitro and in vivo. Nanomedicine (Lond) 7(8):1181–1195. doi:10.2217/nnm.11.177
Mohamed BM, Verma NK, Prina-Mello A, Williams Y, Davies AM, Bakos G, Volkov Y (2011) Activation of stress-related signalling pathway in human cells upon SiO2 nanoparticles exposure as an early indicator of cytotoxicity. J Nanobiotechnol 9:29. doi:10.1186/1477-3155-9-29
Towne DL, Nicholl EE, Comess KM, Galasinski SC, Hajduk PJ, Abraham VC (2012) Development of a high-content screening assay panel to accelerate mechanism of action studies for oncology research. J Biomol Screen 17(8):1005–1017. doi:10.1177/1087057112450050
Cooper SEJ, Kennedy NP, Mohamed BM, Abuzakouk M, Dunne J, Byrne G, Feighery CF (2013) Immunological indicators of coeliac disease activity are not altered by long-term oats challenge. Clin Exp Immunol 171(3):313–318. doi:10.1111/cei.12014
Mohamed BM, Feighery C, Williams Y, Davies A, Kelleher D, Volkov Y, Kelly J, Abuzakouk M (2008) The use of Cellomics to study enterocytes cytoskeletal proteins in coeliac disease patients. Cent Eur J Biol 3(3):258–267
Zock J (2009) Applications of high content screening in life science research. Comb Chem High Throughput Screen 12(9):870–876. doi:10.2174/138620709789383277
Marsh MN (1992) Mucosal pathology in gluten sensitivity. In: Marsh MN (ed) Coeliac disease. Blackwell Scientific, Oxford, pp 136–191
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Cooper, S.E.J. et al. (2015). Adaptation of a Cell-Based High Content Screening System for the In-Depth Analysis of Celiac Biopsy Tissue. In: Ryan, A. (eds) Celiac Disease. Methods in Molecular Biology, vol 1326. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2839-2_8
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DOI: https://doi.org/10.1007/978-1-4939-2839-2_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2838-5
Online ISBN: 978-1-4939-2839-2
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