title: On-chip selective capture of cancer cells and ultrasensitive fluorescence detection of survivin mRNA in a single living cell.
abstract: The rapid recognition of cancer cells and detection of tumor biomarker survivin mRNA plays a critical role in the early diagnosis of many cancers. Based on the integration of specific cancer cell capture and intracellular survivin mRNA detection, this work presents a novel and sensitive on-chip approach for the bioanalysis of survivin mRNA in a single living cell. The microchannel surface was firstly modified with a prostate stem cell antigen (PSCA) monoclonal antibody as the recognition element for prostate cancer cells (PC-3). As a result of the antigen-antibody specific affinity interactions, PC-3 cells could be selectively captured on the microchannel surface. After cell capture, nano-sized graphene oxide-poly(ethylene glycol) bis(amine) (NGO-PEG) was employed as a quencher and carrier of a signal tag, fluorescein isothiocyanate (FITC)-labeled antisense oligonucleotide (F-S1), which is complementary to part of survivin mRNA (target survivin mRNA), to transfect into the captured PC-3 cells. Upon the selective binding of S1 to intracellular survivin mRNA, F-S1 will be released from the NGO-PEG, inducing the fluorescence recovery of FITC. This antibody-based microfluidic device enables simple and inexpensive monitoring of the amount of survivin mRNA in single captured cell without the need for sample pretreatment. The survivin mRNA content in each PC-3 cell was estimated to be (4.8 ± 1.8) × 10(6) copies. This strategy opens a different perspective for ultrasensitive survivin mRNA detection, which may facilitate the early screening for malignancy.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, P.R.China. xujj@nju.edu.cn.
authors.forename: Xiang-Ling
authors.initials: XL
authors.lastname: Li
authors.forename: Shu
authors.initials: S
authors.lastname: Shan
authors.forename: Meng
authors.initials: M
authors.lastname: Xiong
authors.forename: Xing-Hua
authors.initials: XH
authors.lastname: Xia
authors.forename: Jing-juan
authors.initials: JJ
authors.lastname: Xu
authors.forename: Hong-Yuan
authors.initials: HY
authors.lastname: Chen
mesh_terms: D000588:Amines / Q000737:chemistry; D017209:Apoptosis; D015374:Biosensing Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006108:Graphite / Q000737:chemistry; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D042541:Intracellular Space / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D049329:Nanostructures / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000235:genetics; D011092:Polyethylene Glycols / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D059010:Single-Cell Analysis / Q000379:methods*; D013050:Spectrometry, Fluorescence; D013499:Surface Properties; D000077022:Survivin
chemical_list: D000588:Amines; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D000077022:Survivin; D011092:Polyethylene Glycols; D006108:Graphite; D016650:Fluorescein-5-isothiocyanate