Quantifying immunogold localization patterns on electron microscopic thin sections of placenta: recent developments. 2009

T M Mayhew
Centre for Integrated Systems Biology and Medicine, School of Biomedical Sciences, E Floor, Queen's Medical Centre, University of Nottingham, Nottingham NG7 2UH, UK. terry.mayhew@nottingham.ac.uk

In recent years, there have been important advances in the quantification of high-resolution (electron microscopical) images of tissue sections on which colloidal gold-labelled probes serve to identify and localize interesting target antigens. With these new methods, the distributions of gold particle counts across volume-occupying and/or surface-occupying compartments can be compared within or between experimental groups of cells, tissues or organs. Method I (for within-group comparisons) tests whether or not there is preferential labelling of compartments by comparing observed and expected gold labelling distributions using Chi-squared (chi2) analyses. To this end, estimators of gold labelling intensity (labelling density, LD, and/or relative labelling index, RLI) are used to analyse one category of compartment (volume or surface occupiers) or a mixture of categories (volume and surface occupiers). This involves estimating compartment size taking into account specimen magnification and, in its most efficient form, is achieved simply by counting chance events after superimposing random test probes (lattices of points and/or lines). RLI=1 indicates random labelling but higher RLI values indicate the degree to which a compartment departs from random labelling. Method II (drawing between-group comparisons) does not require information about compartment size or specimen magnification. Comparisons are drawn using the observed gold particle counts in different groups and by combining chi2 analysis with contingency table analysis. Both methods rely on multistage systematic uniform random sampling of specimens and unbiased counting of gold particles associated with different compartments. Statistical degrees of freedom are determined by the numbers of compartments and experimental groups. Together with RLI values, compartmental values of chi2 which contribute substantially to total chi2 identify the principal sites of within- and between-group differences. The methods are illustrated using data taken from studies aimed at localising protein antigens (caveolin-1 and GLUT1) in specimens of term human placenta.

UI MeSH Term Description Entries
D007150 Immunohistochemistry Histochemical localization of immunoreactive substances using labeled antibodies as reagents. Immunocytochemistry,Immunogold Techniques,Immunogold-Silver Techniques,Immunohistocytochemistry,Immunolabeling Techniques,Immunogold Technics,Immunogold-Silver Technics,Immunolabeling Technics,Immunogold Silver Technics,Immunogold Silver Techniques,Immunogold Technic,Immunogold Technique,Immunogold-Silver Technic,Immunogold-Silver Technique,Immunolabeling Technic,Immunolabeling Technique,Technic, Immunogold,Technic, Immunogold-Silver,Technic, Immunolabeling,Technics, Immunogold,Technics, Immunogold-Silver,Technics, Immunolabeling,Technique, Immunogold,Technique, Immunogold-Silver,Technique, Immunolabeling,Techniques, Immunogold,Techniques, Immunogold-Silver,Techniques, Immunolabeling
D008867 Microtomy The technique of using a microtome to cut thin or ultrathin sections of tissues embedded in a supporting substance. The microtome is an instrument that hold a steel, glass or diamond knife in clamps at an angle to the blocks of prepared tissues, which it cuts in sections of equal thickness. Thin Sectioning,Ultramicrotomy,Sectioning, Thin,Sectionings, Thin,Thin Sectionings
D002824 Chorionic Villi Threadlike vascular projections of the chorion. Chorionic villi may be free or embedded within the DECIDUA forming the site for exchange of substances between fetal and maternal blood (PLACENTA). Placental Villi,Labyrinth of the Placenta,Labyrinthine Placenta,Placental Labyrinth Layer,Chorionic Villus,Labyrinth Layer, Placental,Placental Labyrinth Layers,Placental Villus,Villi, Chorionic,Villi, Placental,Villus, Chorionic,Villus, Placental
D004730 Endothelium, Vascular Single pavement layer of cells which line the luminal surface of the entire vascular system and regulate the transport of macromolecules and blood components. Capillary Endothelium,Vascular Endothelium,Capillary Endotheliums,Endothelium, Capillary,Endotheliums, Capillary,Endotheliums, Vascular,Vascular Endotheliums
D005260 Female Females
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D014327 Trophoblasts Cells lining the outside of the BLASTOCYST. After binding to the ENDOMETRIUM, trophoblasts develop into two distinct layers, an inner layer of mononuclear cytotrophoblasts and an outer layer of continuous multinuclear cytoplasm, the syncytiotrophoblasts, which form the early fetal-maternal interface (PLACENTA). Cytotrophoblasts,Syncytiotrophoblasts,Trophoblast,Cytotrophoblast,Syncytiotrophoblast
D016253 Microscopy, Immunoelectron Microscopy in which the samples are first stained immunocytochemically and then examined using an electron microscope. Immunoelectron microscopy is used extensively in diagnostic virology as part of very sensitive immunoassays. Immunoelectron Microscopy,Microscopy, Immuno-Electron,Immuno-Electron Microscopies,Immuno-Electron Microscopy,Immunoelectron Microscopies,Microscopies, Immuno-Electron,Microscopies, Immunoelectron,Microscopy, Immuno Electron
D051242 Caveolin 1 A tyrosine phosphoprotein that plays an essential role in CAVEOLAE formation. It binds CHOLESTEROL and is involved in LIPIDS transport, membrane traffic, and SIGNAL TRANSDUCTION. Caveolin-1,VIP21 Protein,Vesicular Integral Membrane Protein 21 kDa,alpha-Caveolin,beta-Caveolin,alpha Caveolin,beta Caveolin
D051272 Glucose Transporter Type 1 A ubiquitously expressed glucose transporter that is important for constitutive, basal GLUCOSE transport. It is predominately expressed in ENDOTHELIAL CELLS and ERYTHROCYTES at the BLOOD-BRAIN BARRIER and is responsible for GLUCOSE entry into the BRAIN. Erythrocyte Glucose Transporter,GLUT-1 Protein,GLUT1 Protein,SLC2A1 Protein,Solute Carrier Family 2, Facilitated Glucose Transporter, Member 1 Protein,GLUT 1 Protein,Glucose Transporter, Erythrocyte

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