title: Comparison of optical data from flow cytometry and microscopy of leukocytes after exposure to specific reagents.
abstract: Flow cytometry and microscopy are equally important in cell analysis. However, few reports have compared the optical data (cell size, internal complexity and fluorescent signal) from flow cytometry and microscopy. In this study, we compared the scattergram from XN-series, a flow cytometry based hematology analyzer with microscopic images of similarly treated leukocytes, and investigated the correlation between the appearance in the scattergram and cell size, internal complexity and fluorescence intensity. Healthy human peripheral blood was analyzed using the XN analyzer. For microscopic comparison, five types of leukocytes (monocytes, lymphocytes, basophils, neutrophils and eosinophils) were isolated from the peripheral blood by centrifugation and magnetic cell sorting, treated with a specific reagent and analyzed using electron microscopy, laser microscopy and confocal laser microscopy. Cell size, residual internal structures and fluorescence intensity correlated with intensity of forward-scattering, side scattering and fluorescent light. In this study, optical data from a clinically used hematology analyzer was clarified using microscopic images.
authors.affiliation: Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Takagi
authors.affiliation: Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan kono.mari@sysmex.co.jp.
authors.forename: Mari
authors.initials: M
authors.lastname: Kono
authors.affiliation: Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan.
authors.forename: Shiori
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Wada
authors.affiliation: Scientific Research, Scientific Affairs, Sysmex Corporation, 1-3-2 Murotani, Nishi-ku, Kobe 651-2241, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Morikawa
mesh_terms: D001491:Basophils / Q000187:drug effects / Q000648:ultrastructure; D048429:Cell Size / Q000187:drug effects; D004804:Eosinophils / Q000187:drug effects / Q000648:ultrastructure; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D007962:Leukocytes / Q000187:drug effects / Q000648:ultrastructure*; D008214:Lymphocytes / Q000187:drug effects / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000187:drug effects / Q000648:ultrastructure; D009504:Neutrophils / Q000187:drug effects / Q000648:ultrastructure
chemical_list: D005456:Fluorescent Dyes
keywords: electron microscopy; flow cytometry; fluorescent intensity; fluorescent microscopy; leukocytes; scatter light intensity