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Gray in the experimental determination of the significance of medical research and application

- The use of an image pixel in the spatial representation; (2) to quantify - the degree of brightness of each pixel, said discrete values, called gray. Image (i, j) position of the gray value of pixel f (i, j), is the pixel intensity (Intensity) of the description. Gray-scale image analysis system determined by the system design, the brightness of a pixel with a single byte of information stored when 2. = 256 gray levels, with double-byte storage when 2 = 65,536 shades of gray. The brightness of the image analysis system to provide measurement parameters are: average gray value (AverageGrayValue) and total gray values (TotalGrayValue). Average gray value of the AGV = ΣGrayValue / NumberofPixel (gray divided by the total number of pixels constitute the object under test). Based on the above theory, the average gray value of medical research in experimental biological samples can be directly reflected in the brightness of the fluorescence intensity is mainly used in quantitative and quantitative electrophoresis products. 1.1 l fluorescence quantitative immunofluorescence method of quantitative analysis of the current depth development of immunology, as long as the application of antigen-antibody reaction can be with their determination. In receptor localization, quantitative study, using antibodies with a fluorescent marker protein hormone receptors, immune fluorescence of a single protein molecules labeled only the tag, the method is relatively simple. Fluorescent double labeling cells also labeled two protein molecules, usually choose between two fluorescent secondary antibodies brought as far as possible away from the emission spectrum of tangled, such as: FITC (520) and TR1. It (572), FITC (520) and cY5 (667) and so on. Escherichia coli lipopolysaccharide in human peritoneal mesothelial cells NF. Effects of KB, the use of immunofluorescence between the standard HTC / PI double staining to observe the NF. KBP Shu in human peritoneal mesothelial cells and nuclear transfer of the case situation, PI is propidium iodide, fluorescent red cells issued, FITC labeled secondary antibody, stained NF. KB made of green fluorescence. Measuring the average gray value of green fluorescent quantitative NF. KB expression. Normal, P65 expression in the cytoplasm, once activated their entry into the nucleus, when green fluorescent and red fluorescent yellow fluorescence appeared at the same time, indicating that nuclear translocation of P65 activation occurs. Received Date】【2006-07-111.2GFP quantitative gene expression product GFP (green fluorescent protein) of the current through the conversion, there are a variety of different colored fluorescent emission of fluorescent proteins, such as: BFP (Blue), YFP (YeUow), CFP (Cyan), etc., BFP fusion protein and the GFP fusion protein can be double-transfected cells,[link widoczny dla zalogowanych], the multi-color fluorescent protein location. And GFPDNA foreign gene linked to, GFP as reporter gene foreign genes regular monitoring of the expression of foreign genes, the determination of the fluorescence intensity of GFP is quantified by the average gray value of the expression of foreign genes. 1.3 Quantitative fluorescence in situ hybridization in the karyotype analysis and gene mapping, often using fluorescence in situ hybridization (FISH), in situ hybridization, based on different gene probe was labeled by different fluorescence in each species for a specific chromosome. Determined by microscopic fluorescence imaging system that is the average fluorescence intensity of gray scale, quantitative expression of fluorescence in situ hybridization. l. 4, intracellular ion concentrations of quantitative fluorescence image analysis system MetaFluo measured in living cells ions in the external drug the dynamic changes (such as: ca ¨, N0, K, №, Mg plasma concentration change),[link widoczny dla zalogowanych], according to cell type and the increase drug test requirements, determine the conditions for fluorescence image acquisition,[link widoczny dla zalogowanych], real time image acquisition set (ImngStack), lock the cells of interest to measure the average gray value (ie fluorescence intensity),[link widoczny dla zalogowanych], the computer automatically draw the fluorescence intensity time curve, reflecting the relative ion concentration. 2 Quantitative electrophoresis electrophoresis with Western blot technique and the type and breadth of application of the rapid development in recent years has become an important means of biological research,[link widoczny dla zalogowanych], including protein nucleic acid gel electrophoresis studies in the preferred method. Electrophoretic methods are commonly used: cellulose acetate membrane electrophoresis cable, gel electrophoresis (agarose gel AGE; polyacrylamide gel PAGE. SDS. PAGE), electrophoretic transfer blot (Southern, Northern, Western). Workstation using image analysis in electrophoresis support media measurement products FluoChemV2.0 band or the average gray value of G imprinting or integrated optical density IDV (IDV = G × Area). V-gray values were measured directly on behalf of the luminous intensity of the object, as with the integrated optical density values reflect the relative content of stained materials.