Journal: Biology of reproduction

Article Title: Quantified colocalization reveals heterotypic histocompatibility class I antigen associations on trophoblast cell membranes: relevance for human pregnancy.

doi: 10.1095/biolreprod.113.111963

Figure Lengend Snippet: FIG. 1. Colocalization detection and quantification of HLA receptors on the cell surface of human trophoblast-derived choriocarcinoma cells. A) 3D image of a JEG-3 cell showing blue (nucleus), green (HLA-E), red (HLA-C), and yellow/orange potentially colocalized particles. B) Mask image of single cell (gray) shown in A. Cell outline was traced using differential interference contrast and its fluorescence image. C) 3D scatter plot of a single cell depicting the relationship between HLA-E (green asterisks) and HLA-C (red asterisks). Blue circles designate colocalized HLA-E and -C molecules. Total number of red, green, and colocalized spots per cell is quantified. D) Color-coded 3D scatter plot for Euclidean distance map of colocalized particles. The color scale indicates the distance between the centroids of colocalized pairs ranging from 10–165 nm (colocalization distance threshold d¼165 nm). The corresponding histogram represents colocalization distance on the x-axis and the number of colocalizations on the y-axis. Note that the grid for cellular dimensions uses micrometers.

Article Snippet: Briefly, the colocalization algorithm uses MATLAB software (version R2010b with Image Processing Toolbox; MathWorks Inc., Natick, Massachusetts) and identifies punctate 3D blob-like features corresponding to fluorescent particles.

Techniques: Derivative Assay, Fluorescence

Journal: Biology of reproduction

Article Title: Quantified colocalization reveals heterotypic histocompatibility class I antigen associations on trophoblast cell membranes: relevance for human pregnancy.

doi: 10.1095/biolreprod.113.111963

Figure Lengend Snippet: FIG. 3. Localization of HLA-C, -E, and -G on the cell surface of ACH-3P and JEG-3 cells, determined by confocal microscopy analysis. Green and red indicate pairings as follows: HLA-G (green)/HLA-E (red), HLA-G (green)/HLA-C (red), and HLA-C (red)/HLA-E (green). Yellow/orange fluorescence reveals the potential colocalization of two antigens. 3D images were acquired in stacks, with z-direction step size 0.14 lm. Single-plane section of z-stack is shown in three directions as xy, yz, and zx. The cutting planes of the cell section are indicated by gray/pale horizontal and vertical lines. A representative single cell from each HLA group is shown. Bar ¼ 10 lm.

Article Snippet: Briefly, the colocalization algorithm uses MATLAB software (version R2010b with Image Processing Toolbox; MathWorks Inc., Natick, Massachusetts) and identifies punctate 3D blob-like features corresponding to fluorescent particles.

Techniques: Confocal Microscopy, Fluorescence