首页> 外文期刊>Journal of Microscopy >A METHOD TO COMPENSATE FOR LIGHT ATTENUATION WITH DEPTH IN THREE-DIMENSIONAL DNA IMAGE CYTOMETRY USING A CONFOCAL SCANNING LASER MICROSCOPE
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A METHOD TO COMPENSATE FOR LIGHT ATTENUATION WITH DEPTH IN THREE-DIMENSIONAL DNA IMAGE CYTOMETRY USING A CONFOCAL SCANNING LASER MICROSCOPE

机译:共聚焦扫描激光显微镜在三维DNA图像细胞学中深度补偿光衰减的方法

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摘要

A method to compensate for attenuation of detected light with increased depth of the collected optical section, and its application in three-dimensional (3-D) DNA image cytometry is described. The method is based on studying the stack of 2-D histograms that can be formed from each consecutive pair of sections in a stack of optical serial sections. An attenuation factor is calculated interactively and a new compensated section series is computed. Formalin-fixed paraffin-embedded rat tissue was stained with propidium iodide, Each cell nucleus is extracted by thresholding and its total intensity is calculated. The coefficient of variation (CV) of the total intensity of all cells Tn each stack is computed. For comparison the CV of the same cells is computed in the uncompensated stacks. This study shows a significantly lower CV for the compensated data, thus contributing to the accuracy of DNA quantification in 3-D DNA image cytometry. [References: 11]
机译:描述了一种用于补偿随着收集的光学部分深度的增加而增加的检测光衰减的方法,及其在三维(3-D)DNA图像细胞仪中的应用。该方法基于研究二维直方图的堆栈,该堆栈可以由光学序列部分堆栈中的每个连续对部分形成。交互计算衰减因子,并计算新的补偿截面系列。用碘化丙锭对福尔马林固定石蜡包埋的大鼠组织进行染色,通过阈值提取每个细胞核,并计算其总强度。计算每个堆叠的所有单元Tn的总强度的变异系数(CV)。为了比较,在未补偿的堆栈中计算相同单元的CV。这项研究表明,补偿数据的CV明显降低,从而有助于3-D DNA图像细胞术中DNA定量的准确性。 [参考:11]

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