Abstract
Imaging flow cytometry is an emerging imaging technology that combines features of both conventional flow cytometry and fluorescence microscopy allowing quantification of the imaging parameters. The analysis of protein posttranslational modifications by glycosylation using imaging flow cytometry constitutes an important bioimaging tool in the glycobiology field. This technique allows quantification of the glycan fluorescence intensity, co-localization with proteins, and evaluation of the membrane/cytoplasmic expression. In this chapter we provide the guidelines to analyze glycan expression, particularly the β1,6 GlcNAc branched N-glycans, on the membrane of intestinal T cells from inflammatory bowel disease patients.
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Acknowledgments
This work was supported by grants from the Portuguese Foundation for Science and Technology (FCT), project grants (PTDC/DTP-PIC/0560/2014; PTDC/BBB-EBI/0786/2012; EXPL/BIM-MEC/0149/2012), “financiados no âmbito do Programa Operacional Temático Factores de Competitividade (COMPETE) e comparticipado pelo fundo Comunitário Europeu FEDER,” e do Quadro de Referência Estratégia Nacional QREN. This work was further supported by a Portuguese grant from “Grupo de Estudo da Doença Inflamatória Intestinal” (GEDII). This work had also the finantial support of FCT/MEC through National Funds and, when applicable, co-financed by the FEDER via the PT2020 Partnership Agreement under the 4293 Unit I&D. S.S.P. (SFRH/BPD/63094/2009) also acknowledges FCT. A.M.D. PD/BD/105982/2014 also acknowledges FCT and BiotechHealth Doctoral Programme. The Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) integrates the Institute for Research and Innovation in Health (I3S), which is partially supported by the Portuguese Foundation for Science and Technology (FCT).
Data was acquired at the Bioimaging Center for Biomaterials and Regenerative Therapies (b.IMAGE, INEB, Porto, Portugal).
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Dias, A.M., Almeida, C.R., Reis, C.A., Pinho, S.S. (2016). Studying T Cells N-Glycosylation by Imaging Flow Cytometry. In: Barteneva, N., Vorobjev, I. (eds) Imaging Flow Cytometry. Methods in Molecular Biology, vol 1389. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3302-0_11
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DOI: https://doi.org/10.1007/978-1-4939-3302-0_11
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