PMID- 38042143 OWN - NLM STAT- MEDLINE DCOM- 20240105 LR - 20240105 IS - 1873-3573 (Electronic) IS - 0039-9140 (Linking) VI - 269 DP - 2024 Mar 1 TI - Detecting EGFR gene amplification using a fluorescence in situ hybridization platform based on digital microfluidics. PG - 125444 LID - S0039-9140(23)01195-5 [pii] LID - 10.1016/j.talanta.2023.125444 [doi] AB - Signal transduction mediated by epidermal growth factor receptor (EGFR) gene affects the proliferation, invasion, metastasis, and angiogenesis of tumor cells. In particular, non-small cell lung cancer (NSCLC) patients with increased in copy number of EGFR gene are often sensitive to tyrosine kinase inhibitors. Despite being the standard for detecting EGFR amplification in the clinic, fluorescence in situ hybridization (FISH) traditionally involves repetitive and complex benchtop procedures that are not only time consuming but also require well-trained personnel. To address these limitations, we develop a digital microfluidics-based FISH platform (DMF-FISH) that automatically implements FISH operations. This system mainly consists of a DMF chip for reagent operation, a heating array for temperature control and a signal processing system. With the capability of automatic droplet handling and efficient temperature control, DMF-FISH performs cell digestion, gradient elution, hybridization and DAPI staining without manual intervention. In addition to operational feasibility, DMF-FISH yields comparable performance with the benchtop FISH protocol but reducing the consumption of DNA probe by 87 % when tested with cell lines and clinical samples. These results highlight unique advantages of the fully automated DMF-FISH system and thus suggest its great potential for clinical diagnosis and personalized therapy of NSCLC. CI - Copyright (c) 2023. Published by Elsevier B.V. FAU - Shen, Chuanjie AU - Shen C AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing, 100049, China. FAU - Zhan, Cheng AU - Zhan C AD - Department of Thoracic Surgery, Zhongshan Hospital, Fudan University, Shanghai, 200030, China. FAU - Tong, Zhaoduo AU - Tong Z AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing, 100049, China. FAU - Yin, Hao AU - Yin H AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing, 100049, China. FAU - Hui, Jianan AU - Hui J AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China. FAU - Qiu, Shihui AU - Qiu S AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing, 100049, China. FAU - Li, Qiushi AU - Li Q AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China. FAU - Xu, Xin AU - Xu X AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China. FAU - Ma, Hui AU - Ma H AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China. FAU - Wu, Zhenhua AU - Wu Z AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing, 100049, China. FAU - Shi, Nan AU - Shi N AD - Suzhou Inst Nanotech & Nanob, Chinese Academy of Sciences, Suzhou, 215123, China. Electronic address: nshi2021@sinano.ac.cn. FAU - Mao, Hongju AU - Mao H AD - State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai, 200050, China; Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Sciences, Beijing, 100049, China. Electronic address: hjmao@mail.sim.ac.cn. LA - eng PT - Journal Article DEP - 20231125 PL - Netherlands TA - Talanta JT - Talanta JID - 2984816R RN - EC 2.7.10.1 (ErbB Receptors) RN - EC 2.7.10.1 (EGFR protein, human) SB - IM MH - Humans MH - *Carcinoma, Non-Small-Cell Lung/pathology MH - *Lung Neoplasms/pathology MH - Gene Amplification MH - ErbB Receptors/genetics/metabolism MH - In Situ Hybridization, Fluorescence/methods MH - Microfluidics MH - Gene Dosage MH - Mutation OTO - NOTNLM OT - Digital microfluidics OT - EGFR amplification OT - Fluorescence in situ hybridization COIS- Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2023/12/03 00:42 MHDA- 2024/01/05 06:42 CRDT- 2023/12/02 18:13 PHST- 2023/09/12 00:00 [received] PHST- 2023/11/16 00:00 [revised] PHST- 2023/11/18 00:00 [accepted] PHST- 2024/01/05 06:42 [medline] PHST- 2023/12/03 00:42 [pubmed] PHST- 2023/12/02 18:13 [entrez] AID - S0039-9140(23)01195-5 [pii] AID - 10.1016/j.talanta.2023.125444 [doi] PST - ppublish SO - Talanta. 2024 Mar 1;269:125444. doi: 10.1016/j.talanta.2023.125444. Epub 2023 Nov 25.