bright field microscopy as an alternative to whole cell fluorescence in automated analysis of macrophage images亮视场显微镜替代整个细胞荧光巨噬细胞图像的自动分析.pdf
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Bright Field Microscopy as an Alternative to Whole Cell
Fluorescence in Automated Analysis of Macrophage
Images
1,2 1,2 1 1 1
Jyrki Selinummi , Pekka Ruusuvuori , Irina Podolsky , Adrian Ozinsky , Elizabeth Gold , Olli Yli-
2 1 1,2,3,4
Harja , Alan Aderem , Ilya Shmulevich *
1 Institute for Systems Biology, Seattle, Washington, United States of America, 2 Department of Signal Processing, Tampere University of Technology, Tampere, Finland,
3 Department of Bioengineering, University of Washington, Seattle, Washington, United States of America, 4 Department of Electrical Engineering, University of
Washington, Seattle, Washington, United States of America
Abstract
Background: Fluorescence microscopy is the standard tool for detection and analysis of cellular phenomena. This
technique, however, has a number of drawbacks such as the limited number of available fluorescent channels in
microscopes, overlapping excitation and emission spectra of the stains, and phototoxicity.
Methodology: We here present and validate a method to automatically detect cell population outlines directly from bright
field images. By imaging samples with several focus levels forming a bright field z -stack, and by measuring the intensity
variations of this stack over the z -dimension, we construct a new two dimensional projection image of increased contrast.
With additional information for locations of each cell, such as stained nuclei, this bright field projection image can be used
instead of whole cell fluorescence to locate borders of individual cells, separating touching cells, and enabling
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