Multiple Tissue Antigen Analysis by Sequential Immunofluorescence Staining and Multi-dimensional Image Analysis
نویسندگان
چکیده
A novel method for sequential immunofluorescence staining, which, in combination with 3D image registration and segmentation, can be used to increase the number of antigens that can be observed simultaneously in single cells in tissue sections is presented. Visualization of more than one antigen by multicolor immunostaining is often desirable or even necessary, both for quantitative studies and to explore spatial relationships of functional significance. Sequential staining, meaning repeated application and removal of fluorescence markers, greatly increases the number of different antigens that can be visualized and quantified in single cells using digital imaging fluorescence microscopy. Quantification and efficient objective analysis of the image data requires digital image analysis. A method for 3D image registration combined with 2D and 3D segmentation and 4D extraction of data is described.
منابع مشابه
Sequential immunofluorescence staining and image analysis for detection of large numbers of antigens in individual cell nuclei.
BACKGROUND Visualization of more than one antigen by multicolor immunostaining is often desirable or even necessary to explore spatial and temporal relationships of functional significance. Previously presented staining protocols have been limited to the visualization of three or four antigens. METHODS Immunofluorescence staining was performed both on slices of formalin-fixed tissue and on ce...
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