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Imaging Cytometry of Human Leukocytes with Third Harmonic Generation Microscopy.


ABSTRACT: Based on third-harmonic-generation (THG) microscopy and a k-means clustering algorithm, we developed a label-free imaging cytometry method to differentiate and determine the types of human leukocytes. According to the size and average intensity of cells in THG images, in a two-dimensional scatter plot, the neutrophils, monocytes, and lymphocytes in peripheral blood samples from healthy volunteers were clustered into three differentiable groups. Using these features in THG images, we could count the number of each of the three leukocyte types both in vitro and in vivo. The THG imaging-based counting results agreed well with conventional blood count results. In the future, we believe that the combination of this THG microscopy-based imaging cytometry approach with advanced texture analysis of sub-cellular features can differentiate and count more types of blood cells with smaller quantities of blood.

SUBMITTER: Wu CH 

PROVIDER: S-EPMC5109028 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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Imaging Cytometry of Human Leukocytes with Third Harmonic Generation Microscopy.

Wu Cheng-Ham CH   Wang Tzung-Dau TD   Hsieh Chia-Hung CH   Huang Shih-Hung SH   Lin Jong-Wei JW   Hsu Szu-Chun SC   Wu Hau-Tieng HT   Wu Yao-Ming YM   Liu Tzu-Ming TM  

Scientific reports 20161115


Based on third-harmonic-generation (THG) microscopy and a k-means clustering algorithm, we developed a label-free imaging cytometry method to differentiate and determine the types of human leukocytes. According to the size and average intensity of cells in THG images, in a two-dimensional scatter plot, the neutrophils, monocytes, and lymphocytes in peripheral blood samples from healthy volunteers were clustered into three differentiable groups. Using these features in THG images, we could count  ...[more]

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