The sampling limit in fluorescence microscopy

被引:11
作者
Heintzmann, Rainer
Sheppard, Colin J. R.
机构
[1] Kings Coll London, Randall Div Cell & Mol Biophys, London SE1 1UL, England
[2] Natl Univ Singapore, Div Bioengn, Singapore 117576, Singapore
[3] Natl Univ Singapore, Dept Diagnost Radiol, Singapore 119074, Singapore
关键词
fluorescence microscopy; sampling; pixel size; optical transfer function; imaging;
D O I
10.1016/j.micron.2006.07.017
中图分类号
TH742 [显微镜];
学科分类号
摘要
Sampling in fluorescence microscopy is treated using the concept of the three-dimensional (3D) optical transfer function (OTF). The border of the OTF frequency surface defines the required minimum sampling. The shape of the OTF is derived from simple considerations and valid for far-field high numerical aperture, vector theory. Optimal regular sampling is achieved by a hexagonal grid in 2D, and corresponding hexagonal structures, body-centered cubic (bcc) and hexagonal close-packed (hcp) structures, in 3D. As compared to standard (rectilinear grid) sampling a reduction of 13.4% in 2D and 29.3% in 3D can be achieved with optimized sampling. This reduction in data size is also accompanied by an imaging speed improvement, a reduction of sample bleaching, and can lead to imaging with better signal to noise ratio. (c) 2006 Elsevier Ltd. All rights reserved.
引用
收藏
页码:145 / 149
页数:5
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