Laser scanning cytometry: capturing the immune system in situ

McGrath, M. A., Morton, A. M. and Harnett, M. M. (2011) Laser scanning cytometry: capturing the immune system in situ. Methods in Cell Biology, 102, pp. 231-260. (doi: 10.1016/B978-0-12-374912-3.00009-2)

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Until recently, it has not been possible to image and functionally correlate the key molecular and cellular events underpinning immunity and tolerance in the intact immune system. Certainly, the field has been revolutionized by the advent of tetramers to identify physiologically relevant specificities of T cells, and the introduction of models in which transgenic T-cell receptor and/or B-cell receptor-bearing lymphocytes are adoptively transferred into normal mice and can then be identified by clonotype-specific antibodies using flow cytometry in vitro, or immunohistochemistry ex vivo. However, these approaches do not allow for quantitative analysis of the precise anatomical, phenotypic, signaling, and functional parameters required for dissecting the development of immune responses in health and disease in vivo. Traditionally, assessment of signal transduction pathways has required biochemical or molecular biological analysis of isolated and highly purified subsets of immune system cells. Inevitably, this creates potential artifacts and does not allow identification of the key signaling events for individual cells present in their microenvironment in situ. These difficulties have now been overcome by new methodologies in cell signaling analysis that are sufficiently sensitive to detect signaling events occurring in individual cells in situ and the development of technologies such as laser scanning cytometry that provide the tools to analyze physiologically relevant interactions between molecules and cells of the innate and the adaptive immune system within their natural environmental niche in vivo.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Harnett, Professor Margaret and Morton, Dr Angela and McGrath, Miss Mairi
Authors: McGrath, M. A., Morton, A. M., and Harnett, M. M.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
Journal Name:Methods in Cell Biology
Published Online:24 June 2011
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
271791Dissection of the Cellular and Molecular Basis of Immunoregulation in VivoMargaret HarnettMedical Research Council (MRC)G9901386Infection Immunity and Inflammation Medicine