Validation of an on-chip p16ink4a/Ki-67 dual immunostaining cervical cytology system using microfluidic device technology

• Published in: Scientific reports Vol. 13; no. 1; p. 17052
• Main Authors: Hashimoto, Kei, Kumagai, Tomoo, Nomura, Kyosuke, Miyagawa, Yuko, Tago, Saori, Takasaki, Kazuki, Takahashi, Yuko, Nishida, Haruka, Ichinose, Takayuki, Hirano, Mana, Hiraike, Haruko, Wada-Hiraike, Osamu, Sasajima, Yuko, Kim, Soo Hyeon, Nagasaka, Kazunori
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 10.10.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 631/61; 631/67; 631/67/1517; 639/166/985; 692/53; Cellular biology; Cervical cancer; Cytology; Human papillomavirus; Humanities and Social Sciences; INK4a protein; Lesions; Microfluidics; multidisciplinary; p16 Protein; Science; Science (multidisciplinary); Thin films
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-023-44273-6

More specific screening systems for cervical cancer may become necessary as the human papillomavirus (HPV) vaccine becomes more widespread. Although p16/Ki-67 dual-staining cytology has several advantages, it requires advanced diagnostic skills. Here, we developed an automated on-chip immunostaining method using a microfluidic device. An electroactive microwell array (EMA) microfluidic device with patterned thin-film electrodes at the bottom of each microwell was used for single-cell capture by dielectrophoresis. Immunostaining and dual staining for p16/Ki-67 were performed on diagnosed liquid cytology samples using the EMA device. The numbers of p16/Ki-67 dual-stained cells captured by the EMA device were determined and compared among the cervical intraepithelial neoplasia (CIN) lesion samples. Seven normal, fifteen CIN grade 3, and seven CIN grade 2 samples were examined. The percentage of dual-positive cells was 18.6% in the CIN grade 2 samples and 23.6% in the CIN grade 3 samples. The percentages of dual-positive staining increased significantly as the severity of the cervical lesions increased. p16/Ki67 dual immunostaining using the EMA device is as sensitive as the conventional method of confirming the histopathological diagnosis of cervical samples. This system enables a quantified parallel analysis at the individual cell level.