title: 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.
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Cheng-Ham
authors.initials: CH
authors.lastname: Wu
authors.affiliation: Cardiovascular Center and Division of Cardiology, Department of Internal Medicine, National Taiwan University Hospital and College of Medicine, Taipei 10002, Taiwan.
authors.forename: Tzung-Dau
authors.initials: TD
authors.lastname: Wang
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Chia-Hung
authors.initials: CH
authors.lastname: Hsieh
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Shih-Hung
authors.initials: SH
authors.lastname: Huang
authors.affiliation: Cardiovascular Center and Division of Cardiology, Department of Internal Medicine, National Taiwan University Hospital and College of Medicine, Taipei 10002, Taiwan.
authors.forename: Jong-Wei
authors.initials: JW
authors.lastname: Lin
authors.affiliation: Department of Laboratory Medicine, National Taiwan University Hospital, Taipei 10002, Taiwan.
authors.forename: Szu-Chun
authors.initials: SC
authors.lastname: Hsu
authors.affiliation: Department of Mathematics, University of Toronto, Toronto, Canada.
authors.forename: Hau-Tieng
authors.initials: HT
authors.lastname: Wu
authors.affiliation: Department of Surgery, National Taiwan University Hospital and College of Medicine, Taipei 10002, Taiwan.
authors.forename: Yao-Ming
authors.initials: YM
authors.lastname: Wu
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei 10617, Taiwan.|Molecular Imaging Center, National Taiwan University, Taipei 10617, Taiwan.|Faculty of Health Sciences, University of Macau, Macao SAR, China.
authors.forename: Tzu-Ming
authors.initials: TM
authors.lastname: Liu
mesh_terms: D006801:Humans; D019044:Image Cytometry / Q000295:instrumentation / Q000379:methods*; D007958:Leukocyte Count / Q000295:instrumentation / Q000379:methods; D007962:Leukocytes / Q000166:cytology*; D000073798:Second Harmonic Generation Microscopy / Q000295:instrumentation / Q000379:methods*