Development of on-chip multi-imaging flow cytometry for identification of imaging biomarkers of clustered circulating tumor cells

Hyonchol Kim, Hideyuki Terazono, Yoshiyasu Nakamura, Kazuko Sakai, Akihiro Hattori, Masao Odaka, Mathias Girault, Tokuzo Arao, Kazuto Nishio, Yohei Miyagi, Kenji Yasuda

Research output: Contribution to journalArticle

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Abstract

An on-chip multi-imaging flow cytometry system has been developed to obtain morphometric parameters of cell clusters such as cell number, perimeter, total cross-sectional area, number of nuclei and size of clusters as "imaging biomarkers", with simultaneous acquisition and analysis of both bright-field (BF) and fluorescent (FL) images at 200 frames per second (fps); by using this system, we examined the effectiveness of using imaging biomarkers for the identification of clustered circulating tumor cells (CTCs). Sample blood of rats in which a prostate cancer cell line (MAT-LyLu) had been pre-implanted was applied to a microchannel on a disposable microchip after staining the nuclei using fluorescent dye for their visualization, and the acquired images were measured and compared with those of healthy rats. In terms of the results, clustered cells having (1) cell area larger than 200 μm2 and (2) nucleus area larger than 90 μm2 were specifically observed in cancer cell-implanted blood, but were not observed in healthy rats. In addition, (3) clusters having more than 3 nuclei were specific for cancer-implanted blood and (4) a ratio between the actual perimeter and the perimeter calculated from the obtained area, which reflects a shape distorted from ideal roundness, of less than 0.90 was specific for all clusters having more than 3 nuclei and was also specific for cancer-implanted blood. The collected clusters larger than 300 μm2 were examined by quantitative gene copy number assay, and were identified as being CTCs. These results indicate the usefulness of the imaging biomarkers for characterizing clusters, and all of the four examined imaging biomarkers - cluster area, nuclei area, nuclei number, and ratio of perimeter - can identify clustered CTCs in blood with the same level of preciseness using multi-imaging cytometry.

Original languageEnglish
Article numbere104372
JournalPLoS One
Volume9
Issue number8
DOIs
Publication statusPublished - 2014 Aug 20
Externally publishedYes

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Circulating Neoplastic Cells
Flow cytometry
Biomarkers
flow cytometry
Tumors
biomarkers
Flow Cytometry
Cells
image analysis
Blood
Imaging techniques
blood
Rats
Neoplasms
Gene Dosage
rats
Fluorescent Dyes
cells
gene dosage
Blood Cells

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Development of on-chip multi-imaging flow cytometry for identification of imaging biomarkers of clustered circulating tumor cells. / Kim, Hyonchol; Terazono, Hideyuki; Nakamura, Yoshiyasu; Sakai, Kazuko; Hattori, Akihiro; Odaka, Masao; Girault, Mathias; Arao, Tokuzo; Nishio, Kazuto; Miyagi, Yohei; Yasuda, Kenji.

In: PLoS One, Vol. 9, No. 8, e104372, 20.08.2014.

Research output: Contribution to journalArticle

Kim, Hyonchol ; Terazono, Hideyuki ; Nakamura, Yoshiyasu ; Sakai, Kazuko ; Hattori, Akihiro ; Odaka, Masao ; Girault, Mathias ; Arao, Tokuzo ; Nishio, Kazuto ; Miyagi, Yohei ; Yasuda, Kenji. / Development of on-chip multi-imaging flow cytometry for identification of imaging biomarkers of clustered circulating tumor cells. In: PLoS One. 2014 ; Vol. 9, No. 8.
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