title: A fast and reliable readout method for quantitative analysis of surface-enhanced Raman scattering nanoprobes on chip surface.
abstract: Surface-enhanced Raman scattering techniques have been widely used for bioanalysis due to its high sensitivity and multiplex capacity. However, the point-scanning method using a micro-Raman system, which is the most common method in the literature, has a disadvantage of extremely long measurement time for on-chip immunoassay adopting a large chip area of approximately 1-mm scale and confocal beam point of ca. 1-μm size. Alternative methods such as sampled spot scan with high confocality and large-area scan method with enlarged field of view and low confocality have been utilized in order to minimize the measurement time practically. In this study, we analyzed the two methods in respect of signal-to-noise ratio and sampling-led signal fluctuations to obtain insights into a fast and reliable readout strategy. On this basis, we proposed a methodology for fast and reliable quantitative measurement of the whole chip area. The proposed method adopted a raster scan covering a full area of 100 μm × 100 μm region as a proof-of-concept experiment while accumulating signals in the CCD detector for single spectrum per frame. One single scan with 10 s over 100 μm × 100 μm area yielded much higher sensitivity compared to sampled spot scanning measurements and no signal fluctuations attributed to sampled spot scan. This readout method is able to serve as one of key technologies that will bring quantitative multiplexed detection and analysis into practice.
authors.affiliation: Department of Chemistry Education, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Hyejin
authors.initials: H
authors.lastname: Chang
authors.affiliation: Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Homan
authors.initials: H
authors.lastname: Kang
authors.affiliation: Department of Chemistry Education, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Sinyoung
authors.initials: S
authors.lastname: Jeong
authors.affiliation: Department of Chemistry Education, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Eunbyeol
authors.initials: E
authors.lastname: Ko
authors.affiliation: Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Yoon-Sik
authors.initials: YS
authors.lastname: Lee
authors.affiliation: Department of Nuclear Medicine, Seoul National University Bundang Hospital, Seongnam 463-707, South Korea.
authors.forename: Ho-Young
authors.initials: HY
authors.lastname: Lee
authors.affiliation: Department of Chemistry Education, Seoul National University, Seoul 151-742, South Korea.
authors.forename: Dae Hong
authors.initials: DH
authors.lastname: Jeong
mesh_terms: D007118:Immunoassay / Q000295:instrumentation; D049329:Nanostructures / Q000737:chemistry*; D059629:Signal-To-Noise Ratio; D013059:Spectrum Analysis, Raman / Q000379:methods*