PMID- 25770459 OWN - NLM STAT- MEDLINE DCOM- 20160212 LR - 20150526 IS - 1873-4235 (Electronic) IS - 0956-5663 (Linking) VI - 69 DP - 2015 Jul 15 TI - A fluorescence in situ hybridization (FISH) microfluidic platform for detection of HER2 amplification in cancer cells. PG - 272-9 LID - S0956-5663(15)00145-1 [pii] LID - 10.1016/j.bios.2015.03.003 [doi] AB - Over-expression/amplification of human epidermal growth factor receptors 2 (HER2) is a verified therapeutic biomarker for breast and gastric cancers. HER2 is also served as prognostic biomarker for gastric cancer because HER2 over-expression is associated with a 5-10% increase in cancer related death of gastric cancer. Cancer patients exhibiting HER2 over-expression can significantly improve their overall survival rates by taking the targeting drug Herceptin, which directly targets HER2. However, Herceptin has limited functions toward patients without HER2 over-expression and therefore it needs a highly specific and accurate detection method for diagnosis of HER2 over-expression. Currently, fluorescence in situ hybridization (FISH) technique is routinely employed to detect HER2 amplification. However, it is a labor-intensive, time-consuming hybridization process and is relatively costly. Furthermore, well-trained personnel are required to operate the delicate and complicate process. More importantly, it may take 1-2 days for well-trained personnel to perform a whole FISH assay. Given these limitations, we developed a new, integrated microfluidic FISH system capable of automating the entire FISH protocol which could be performed within a shorter period of time when compared to traditional methods. The microfluidic FISH chip consisted of a microfluidic control module for transportation of small amounts of fluids and a hybridization module to perform the hybridization of DNA probes and cells/tissue samples. With this approach, the new microfluidic chip was capable of performing the whole FISH assay within 20h. Four cell lines, two for non-HER2 amplification and two for HER2 amplification, and two clinical tissue samples, one for non-HER2 amplification and another for HER2 amplification, were used for verifications of the developed chip. Experimental data showed that there was no significant difference between the benchtop protocol and the chip-based protocol. Furthermore, the reagent consumption was greatly reduced ( approximately 70% reduction). Especially, only 2-mul usage for FISH deoxyribonucleic acid (DNA) probe was used, which is five-fold reduction when compared with the traditional method. It is the first time that the entire FISH assay could be automated on a single chip by using tissue samples. The microfluidic system developed herein is therefore promising for rapid, automatic diagnosis of HER2-related diseases by detecting the HER2 gene with minimal consumption of samples and reagents and has a great potential for future pharmacogenetic diagnostics and therapy. CI - Copyright (c) 2015 Elsevier B.V. All rights reserved. FAU - Kao, Kai-Jie AU - Kao KJ AD - Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan. FAU - Tai, Chien-Hsuan AU - Tai CH AD - Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan. FAU - Chang, Wen-Hsin AU - Chang WH AD - Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan. FAU - Yeh, Ta-Sen AU - Yeh TS AD - Department of Surgery, Chang Gung Memorial Hospital at Linkou, Chang Gung University, Taoyuan 333, Taiwan. FAU - Chen, Tse-Ching AU - Chen TC AD - Department of Pathology, Chang Gung Memorial Hospital at Linkou, Chang Gung University, Taoyuan 333, Taiwan. FAU - Lee, Gwo-Bin AU - Lee GB AD - Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan; Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan; Institute of NanoEngineering and Microsystems, National Tsing Hua University, Hsinchu 300, Taiwan. Electronic address: gwobin@pme.nthu.edu.tw. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150303 PL - England TA - Biosens Bioelectron JT - Biosensors & bioelectronics JID - 9001289 RN - 0 (Biomarkers, Tumor) RN - EC 2.7.10.1 (ERBB2 protein, human) RN - EC 2.7.10.1 (Receptor, ErbB-2) SB - IM MH - Biomarkers, Tumor/*metabolism MH - Cell Line, Tumor MH - Equipment Design MH - Equipment Failure Analysis MH - Gene Expression Profiling/*instrumentation MH - Humans MH - In Situ Hybridization, Fluorescence/*instrumentation MH - *Lab-On-A-Chip Devices MH - Neoplasms, Experimental/*metabolism MH - Receptor, ErbB-2/*metabolism MH - Reproducibility of Results MH - Sensitivity and Specificity MH - Up-Regulation OTO - NOTNLM OT - Biomarker OT - Cancer OT - Fluorescence in situ hybridization (FISH) OT - HER2 OT - Herceptin OT - Microfluidics EDAT- 2015/03/17 06:00 MHDA- 2016/02/13 06:00 CRDT- 2015/03/16 06:00 PHST- 2015/01/29 00:00 [received] PHST- 2015/03/02 00:00 [accepted] PHST- 2015/03/16 06:00 [entrez] PHST- 2015/03/17 06:00 [pubmed] PHST- 2016/02/13 06:00 [medline] AID - S0956-5663(15)00145-1 [pii] AID - 10.1016/j.bios.2015.03.003 [doi] PST - ppublish SO - Biosens Bioelectron. 2015 Jul 15;69:272-9. doi: 10.1016/j.bios.2015.03.003. Epub 2015 Mar 3.