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Quantitative phase imaging of cells in a flow cytometry arrangement utilizing Michelson interferometer-based off-axis digital holographic microscopy
Min, Junwei1,2; Yao, Baoli2; Trendafilova, Veselina1; Ketelhut, Steffi1; Kastl, Lena1; Greve, Burkhard3; Kemper, Bjoern1
Department瞬态光学技术国家重点实验室
2019-09
Source PublicationJOURNAL OF BIOPHOTONICS
ISSN1864-063X;1864-0648
Volume12Issue:9
Contribution Rank1
Abstract

We combined Michelson-interferometer-based off-axis digital holographic microscopy (DHM) with a common flow cytometry (FCM) arrangement. Utilizing object recognition procedures and holographic autofocusing during the numerical reconstruction of the acquired off-axis holograms, sharply focused quantitative phase images of suspended cells in flow were retrieved without labeling, from which biophysical cellular features of distinct cells, such as cell radius, refractive index and dry mass, can be subsequently retrieved in an automated manner. The performance of the proposed concept was first characterized by investigations on microspheres that were utilized as test standards. Then, we analyzed two types of pancreatic tumor cells with different morphology to further verify the applicability of the proposed method for quantitative live cell imaging. The retrieved biophysical datasets from cells in flow are found in good agreement with results from comparative investigations with previously developed DHM methods under static conditions, which demonstrates the effectiveness and reliability of our approach. Our results contribute to the establishment of DHM in imaging FCM and prospect to broaden the application spectrum of FCM by providing complementary quantitative imaging as well as additional biophysical cell parameters which are not accessible in current high-throughput FCM measurements.

Keywordautofocusing digital holographic microscopy flow cytometry image flow cytometry label-free cytometry quantitative microscopy quantitative phase imaging
DOI10.1002/jbio.201900085
Indexed BySCI
Language英语
WOS IDWOS:000486778200017
PublisherWILEY-V C H VERLAG GMBH
Citation statistics
Document Type期刊论文
Identifierhttp://ir.opt.ac.cn/handle/181661/31882
Collection瞬态光学技术国家重点实验室
Corresponding AuthorKemper, Bjoern
Affiliation1.Univ Munster, Biomed Technol Ctr, Mendelstr 17, D-48149 Munster, Germany
2.Chinese Acad Sci, Xian Inst Opt & Precis Mech, State Key Lab Transient Opt & Photon, Xian, Shaanxi, Peoples R China
3.Univ Hosp Muenster, Dept Radiotherapy Radiooncol, Munster, Germany
Recommended Citation
GB/T 7714
Min, Junwei,Yao, Baoli,Trendafilova, Veselina,et al. Quantitative phase imaging of cells in a flow cytometry arrangement utilizing Michelson interferometer-based off-axis digital holographic microscopy[J]. JOURNAL OF BIOPHOTONICS,2019,12(9).
APA Min, Junwei.,Yao, Baoli.,Trendafilova, Veselina.,Ketelhut, Steffi.,Kastl, Lena.,...&Kemper, Bjoern.(2019).Quantitative phase imaging of cells in a flow cytometry arrangement utilizing Michelson interferometer-based off-axis digital holographic microscopy.JOURNAL OF BIOPHOTONICS,12(9).
MLA Min, Junwei,et al."Quantitative phase imaging of cells in a flow cytometry arrangement utilizing Michelson interferometer-based off-axis digital holographic microscopy".JOURNAL OF BIOPHOTONICS 12.9(2019).
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