Identification of cells using morphological information of bright field/fluorescent multi-imaging flow cytometer images

Akihiro Hattori, Hyonchol Kim, Hideyuki Terazono, Masao Odaka, Mathias Girault, Kenji Matsuura, Kenji Yasuda

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We have examined the ability of real-time simultaneous measurement of bright field/fluorescent images of cells in an on-chip bright field/fluorescent multi-imaging flow cytometer system. The system consists of (1) a disposable microfluidic hydrofocusing flow cytometry chip, (2) an optical microscopy module with splittable bright field/fluorescent multi-imaging optics, and (3) a real-time image-processing module with a 200 images/s high-speed digital camera. In the double "Y" shape three-way-inlet microfluidic pathways fabricated in the poly(dimethylsiloxane) (PDMS) microchip, we applied fluorescent polystyrene standard beads and HeLa cells stained with fluorescent dye, Hoechst 33258, and measured the z-axis (depth) dependence of the morphological index; the intensity profile of cells and nuclei. Then, we measured the tendency of the blur of bright field/fluorescent images in the simultaneous measurement of bright field/fluorescent images on a single light-receiving surface, and found that their blurs were similar within the same range of the depth of the microfluidic pathway for small cell cluster measurement, 25μm. Hence, the fluorescent images were applied as supporting information of the bright field images of cell clusters at the focal plane for the cell number counting. The result indicates the potential of precise identification of various types of cells by simultaneous morphological analysis of bright field and fluorescent images distributed with a single camera in a wider depth of microfluidic chip as a substitute for conventional biomarker detection.

Original languageEnglish
Article number06JL03
JournalJapanese Journal of Applied Physics
Volume53
Issue number6 SPEC. ISSUE
DOIs
Publication statusPublished - 2014
Externally publishedYes

Fingerprint

Microfluidics
Imaging techniques
cells
chips
Flow cytometry
High speed cameras
Digital cameras
Biomarkers
Polydimethylsiloxane
morphological indexes
Optical microscopy
modules
Optics
Polystyrenes
Image processing
Dyes
Cameras
cytometry
high speed cameras
digital cameras

ASJC Scopus subject areas

  • Engineering(all)
  • Physics and Astronomy(all)

Cite this

Identification of cells using morphological information of bright field/fluorescent multi-imaging flow cytometer images. / Hattori, Akihiro; Kim, Hyonchol; Terazono, Hideyuki; Odaka, Masao; Girault, Mathias; Matsuura, Kenji; Yasuda, Kenji.

In: Japanese Journal of Applied Physics, Vol. 53, No. 6 SPEC. ISSUE, 06JL03, 2014.

Research output: Contribution to journalArticle

@article{0a66eb9e5233497e9af1eae9a2ea350a,
title = "Identification of cells using morphological information of bright field/fluorescent multi-imaging flow cytometer images",
abstract = "We have examined the ability of real-time simultaneous measurement of bright field/fluorescent images of cells in an on-chip bright field/fluorescent multi-imaging flow cytometer system. The system consists of (1) a disposable microfluidic hydrofocusing flow cytometry chip, (2) an optical microscopy module with splittable bright field/fluorescent multi-imaging optics, and (3) a real-time image-processing module with a 200 images/s high-speed digital camera. In the double {"}Y{"} shape three-way-inlet microfluidic pathways fabricated in the poly(dimethylsiloxane) (PDMS) microchip, we applied fluorescent polystyrene standard beads and HeLa cells stained with fluorescent dye, Hoechst 33258, and measured the z-axis (depth) dependence of the morphological index; the intensity profile of cells and nuclei. Then, we measured the tendency of the blur of bright field/fluorescent images in the simultaneous measurement of bright field/fluorescent images on a single light-receiving surface, and found that their blurs were similar within the same range of the depth of the microfluidic pathway for small cell cluster measurement, 25μm. Hence, the fluorescent images were applied as supporting information of the bright field images of cell clusters at the focal plane for the cell number counting. The result indicates the potential of precise identification of various types of cells by simultaneous morphological analysis of bright field and fluorescent images distributed with a single camera in a wider depth of microfluidic chip as a substitute for conventional biomarker detection.",
author = "Akihiro Hattori and Hyonchol Kim and Hideyuki Terazono and Masao Odaka and Mathias Girault and Kenji Matsuura and Kenji Yasuda",
year = "2014",
doi = "10.7567/JJAP.53.06JL03",
language = "English",
volume = "53",
journal = "Japanese Journal of Applied Physics, Part 1: Regular Papers & Short Notes",
issn = "0021-4922",
publisher = "Japan Society of Applied Physics",
number = "6 SPEC. ISSUE",

}

TY - JOUR

T1 - Identification of cells using morphological information of bright field/fluorescent multi-imaging flow cytometer images

AU - Hattori, Akihiro

AU - Kim, Hyonchol

AU - Terazono, Hideyuki

AU - Odaka, Masao

AU - Girault, Mathias

AU - Matsuura, Kenji

AU - Yasuda, Kenji

PY - 2014

Y1 - 2014

N2 - We have examined the ability of real-time simultaneous measurement of bright field/fluorescent images of cells in an on-chip bright field/fluorescent multi-imaging flow cytometer system. The system consists of (1) a disposable microfluidic hydrofocusing flow cytometry chip, (2) an optical microscopy module with splittable bright field/fluorescent multi-imaging optics, and (3) a real-time image-processing module with a 200 images/s high-speed digital camera. In the double "Y" shape three-way-inlet microfluidic pathways fabricated in the poly(dimethylsiloxane) (PDMS) microchip, we applied fluorescent polystyrene standard beads and HeLa cells stained with fluorescent dye, Hoechst 33258, and measured the z-axis (depth) dependence of the morphological index; the intensity profile of cells and nuclei. Then, we measured the tendency of the blur of bright field/fluorescent images in the simultaneous measurement of bright field/fluorescent images on a single light-receiving surface, and found that their blurs were similar within the same range of the depth of the microfluidic pathway for small cell cluster measurement, 25μm. Hence, the fluorescent images were applied as supporting information of the bright field images of cell clusters at the focal plane for the cell number counting. The result indicates the potential of precise identification of various types of cells by simultaneous morphological analysis of bright field and fluorescent images distributed with a single camera in a wider depth of microfluidic chip as a substitute for conventional biomarker detection.

AB - We have examined the ability of real-time simultaneous measurement of bright field/fluorescent images of cells in an on-chip bright field/fluorescent multi-imaging flow cytometer system. The system consists of (1) a disposable microfluidic hydrofocusing flow cytometry chip, (2) an optical microscopy module with splittable bright field/fluorescent multi-imaging optics, and (3) a real-time image-processing module with a 200 images/s high-speed digital camera. In the double "Y" shape three-way-inlet microfluidic pathways fabricated in the poly(dimethylsiloxane) (PDMS) microchip, we applied fluorescent polystyrene standard beads and HeLa cells stained with fluorescent dye, Hoechst 33258, and measured the z-axis (depth) dependence of the morphological index; the intensity profile of cells and nuclei. Then, we measured the tendency of the blur of bright field/fluorescent images in the simultaneous measurement of bright field/fluorescent images on a single light-receiving surface, and found that their blurs were similar within the same range of the depth of the microfluidic pathway for small cell cluster measurement, 25μm. Hence, the fluorescent images were applied as supporting information of the bright field images of cell clusters at the focal plane for the cell number counting. The result indicates the potential of precise identification of various types of cells by simultaneous morphological analysis of bright field and fluorescent images distributed with a single camera in a wider depth of microfluidic chip as a substitute for conventional biomarker detection.

UR - http://www.scopus.com/inward/record.url?scp=84903269939&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84903269939&partnerID=8YFLogxK

U2 - 10.7567/JJAP.53.06JL03

DO - 10.7567/JJAP.53.06JL03

M3 - Article

VL - 53

JO - Japanese Journal of Applied Physics, Part 1: Regular Papers & Short Notes

JF - Japanese Journal of Applied Physics, Part 1: Regular Papers & Short Notes

SN - 0021-4922

IS - 6 SPEC. ISSUE

M1 - 06JL03

ER -