title: A Morphological identification cell cytotoxicity assay using cytoplasm-localized fluorescent probe (CLFP) to distinguish living and dead cells.
abstract: Cell cytotoxicity assays include cell activity assays and morphological identification assays. Currently, all frequently used cytotoxicity assays belong to cell activity assays but suffer from detection limitations. Morphological identification of cell death remains as the gold standard, although the method is difficult to scale up. At present there is no generally accepted morphological identification based cell cytotoxicity assay. In this study, we applied previous developed cell cytoplasm-localized fluorescent probe (CLFP) to display cell morphologies. Under fluorescence microscopy, the fluorescence morphology and intensity of living cells are distinct from dead cells. Based on these characters we extracted the images of living cells from series of samples via computational analysis. Thus, a novel cell morphological identification cytotoxicity assay (CLFP assay) is developed. The performance of the CLFP assay was similar to cell activity assay (MTT assay), but the accuracy of the CLFP assay was superior when measuring the cytotoxicity of active compounds.
authors.affiliation: State Key Laboratory of Bioactive Substances and Function of Natural Medicine, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100050, China.
authors.forename: Fangfang
authors.initials: F
authors.lastname: Lai
authors.affiliation: Department of Applied Mathematics, University of Science and Technology Beijing, Beijing, 100083, China.
authors.forename: Zhengwei
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Beijing Key Laboratory of Active Substances Discovery and Drugability Evaluation, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100050, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wen
authors.affiliation: Department of Mathematics, University of Dundee, Dundee, DD1 4HN, UK.
authors.forename: Jialing
authors.initials: J
authors.lastname: Chen
authors.affiliation: Beijing Key Laboratory of Active Substances Discovery and Drugability Evaluation, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100050, China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Mathematics, University of Dundee, Dundee, DD1 4HN, UK.
authors.forename: Ping
authors.initials: P
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Bioactive Substances and Function of Natural Medicine, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100050, China.
authors.forename: Dali
authors.initials: D
authors.lastname: Yin
authors.affiliation: Beijing Key Laboratory of Active Substances Discovery and Drugability Evaluation, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100050, China. Electronic address: hcui@imm.ac.cn.
authors.forename: Huaqing
authors.initials: H
authors.lastname: Cui
authors.affiliation: State Key Laboratory of Bioactive Substances and Function of Natural Medicine, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100050, China. Electronic address: chxg@imm.ac.cn.
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Chen
mesh_terms: D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001681:Biological Assay / Q000379:methods; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects* / Q000502:physiology; D003593:Cytoplasm / Q000737:chemistry; D003603:Cytotoxins / Q000008:administration & dosage*; D005456:Fluorescent Dyes / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods*; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D003603:Cytotoxins; D005456:Fluorescent Dyes
keywords: Cell death; Cell morphological identification; Computational analysis; Cytoplasm-localized fluorescent probe; Cytotoxicity assay