Bossinger J, Miller M J, Vo K P, Geiduschek E P, Xuong N H
J Biol Chem. 1979 Aug 25;254(16):7986-98.
A method for quantitative analysis of complex film density distributions in autoradiograms is described. The method is intended particularly for measuring the distribution of radioactivity among the proteins resolved by two-dimensional gel electrophoresis but should, of course, be suited to analyzing other two dimensional separations. The film density distribution is first digitized by a high speed rotating drum scanner to generate the image data array that is stored on a magnetic disk. Subsequent analysis involves: 1) data averaging, 2) detection of contours and of their locations, 3) splitting of overlapping spots, 4) conversion of film density to radioactive intensity by means of calibration films, and 5) differentiation and integration to measure the total radioactivity contained in the protein which generates a spot in the autoradiogram. The product of the analysis is a numbered contour map and a table listing coordinates and radioactivity content of each resolved spot. Coordinate transformations for comparison and matching of autoradiograms are also described. A set of utility programs print and graph the data at intermediate stages of the analysis in order to facilitate the checking of procedures and programs.
本文描述了一种用于定量分析放射自显影片中复杂胶片密度分布的方法。该方法特别适用于测量二维凝胶电泳分离出的蛋白质之间的放射性分布，但当然也适用于分析其他二维分离。胶片密度分布首先由高速旋转鼓式扫描仪数字化，以生成存储在磁盘上的图像数据阵列。后续分析包括：1）数据平均，2）轮廓及其位置的检测，3）重叠斑点的分割，4）通过校准胶片将胶片密度转换为放射性强度，5）微分和积分以测量在放射自显影片中产生斑点的蛋白质中所含的总放射性。分析的结果是一个编号的轮廓图和一个列出每个解析斑点的坐标和放射性含量的表格。还描述了用于放射自显影片比较和匹配的坐标变换。一组实用程序在分析的中间阶段打印和绘制数据，以便于检查程序和方案。
