Automated sizing of DNA fragments in atomic force microscope images.
Automated sizing of DNA fragments in atomic force microscope images.
复制标题
自动测定原子力显微镜图像中 DNA 片段的大小。
DOI：
10.1007/bf02518867
复制
发表时间：
1998
影响因子：
3.2
通讯作者：
Hoh,JH
中科院分区：
文献类型：
--
作者：
Spisz,TS;Fang,Y;Reeves,RH;Seymour,CK;Bankman,IN;Hoh,JH
目前用于测量原子力显微镜（AFM）图像中DNA片段长度的技术需要用户操作交互式软件并执行繁琐的易于出错的光标选择。提出了一种算法，它提供了一种自动化的方法，用于确定DNA片段长度从AFM图像，而无需从计算机操作员（如光标选择或鼠标点击）的相互作用。该方法利用图像处理技术定制的DNA片段的AFM图像的特性。与基于图像的手动测量相比，自动测量的平均绝对偏差小于1个像素。从计算长度的直方图确定的DNA长度精确到溶液中实际DNA长度的3%以内。对于250个碱基对长的片段，估计精度在17 nm以内，约为总长度的20%。当该算法很容易地分辨出一个图像中仅相差17 nm的片段时，这种精度得到了证实。片段大小高达2000碱基对已被测试和成功的大小。该算法正在开发，作为新的固态DNA尺寸测量技术的一部分，用于基因分型和物理基因组图谱构建等应用。
Current techniques used to measure lengths of DNA fragments in atomic force microscope (AFM) images require a user to operate interactive software and execute tedious error-prone cursor selections. An algorithm is proposed which provides an automated method for determining DNA fragment lengths from AFM images without interaction from the computer operator (e.g. cursor selections or mouse clicks). The approach utilises image processing techniques tailored to characteristics of AFM images of DNA fragments. The automated measurements have a mean absolute deviation of less than 1 pixel when compared to manual image-based measurements. The DNA length determined from the histogram of calculated lengths is accurate to within 3% of the actual DNA length in solution. For fragments that are 250 base-pairs long, the precision is estimated to be within 17 nm, which is about 20% of the total length. This precision was confirmed when the algorithm easily resolved fragments in one image that differed by only 17 nm. Fragment sizes up to 2000 base-pairs have been tested and successfully sized. This algorithm is being developed as part of a new solid-state DNA sizing technique for applications such as genotyping and construction of physical genome maps.