Imaging Cytometry of Human Leukocytes with Third Harmonic Generation Microscopy

Cheng Ham Wu, Tzung Dau Wang, Chia Hung Hsieh, Shih Hung Huang, Jong Wei Lin, Szu Chun Hsu, Hau Tieng Wu, Yao Ming Wu, Tzu Ming Liu

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish (US)
Article number37210
JournalScientific reports
StatePublished - Nov 15 2016

ASJC Scopus subject areas

  • General


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