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On-chip immunofluorescence analysis of single cervical cells using an electroactive microwell array with barrier for cervical screening

Authors :
Tomoyuki Fujii
Yutaka Osuga
Kazunori Nagasaka
Teruo Fujii
Osamu Wada-Hiraike
Yoshiko Kawata
Takuya Ayabe
Soo Hyeon Kim
Makoto Takeuchi
Haruko Hiraike
Katsutoshi Oda
Mina Yoshida
Saori Tago
Yuko Miyagawa
Source :
Biomicrofluidics
Publication Year :
2019
Publisher :
AIP Publishing, 2019.

Abstract

Several specific tests for cervical screening have been developed recently, including p16/Ki67 dual immunostaining for diagnosing high-risk human papillomavirus positive squamous intraepithelial lesion in the cervix. However, manual screening of cells in an entire glass slide is currently a standard clinical procedure for quantification and interpretation of immunocytochemical features of the cells. Here, we developed a microfluidic device containing an electroactive microwell array with barriers (EMAB) for highly efficient single-cell trapping followed by on-chip immunofluorescence analysis with minimum loss of the sample. EMAB utilizes patterned electrodes at the bottom of cell-sized microwells to trap single cells using dielectrophoresis (DEP) and cell-holding structures behind the microwells to stabilize the position of trapped cells even without DEP. Using the device, we evaluated the performance of p16/Ki67 dual immunostaining of HeLa cells on the chip. The device shows 98% cell-trapping efficiency as well as 92% cell-holding efficiency against the fixed HeLa cells, and we successfully demonstrated high-efficiency on-chip immunofluorescence analysis with minimal loss of sample. p16/Ki67 dual immunostaining using EMAB may be useful for complementary tests for cervical screening in confirming the histopathological diagnosis.

Details

ISSN :
19321058
Volume :
13
Database :
OpenAIRE
Journal :
Biomicrofluidics
Accession number :
edsair.doi.dedup.....6b6a186b183e31b0a5cce554fa649a32
Full Text :
https://doi.org/10.1063/1.5089796