Flat‐Field Lens Grating Microspectrometer for Spectroscopic Multiplex Bioassay

Gi Beom Kim, Jung‐Woo Park, Jaehun Jeon, Hyejeong Jeong, Eun‐Sil Yu, Ki‐Hun Jeong
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Abstract

Spectroscopic multiplex bioassay allows the rapid and simultaneous detection of various biomarkers in biomedical applications. The high cost and bulky configuration of conventional spectrometers hinder the decentralization of biomedical instruments for point‐of‐care diagnostics. Here a flat‐field lens grating‐based microspectrometer (FLG‐µSPEC) is reported for continuous detection of multi‐target nucleic acid amplification. FLG‐µSPEC features the vertical integration of microslit, integrated flat‐field lens and transmission grating (FLG), concave mirror, and CMOS line sensor. FLG effectively corrects the field curvature by flattening the focal plane of dispersed wavelengths and also facilitates compact module packaging. FLG‐µSPEC module is precisely assembled into a dimension of 10 mm × 16 mm × 10 mm. The average spectral resolution exhibits 3.4 ± 0.9 nm over an operating range of 500 to 950 nm. A spectral detection system utilizing FLG‐µSPEC demonstrates the amplification of lambda DNA (𝜆‐DNA) for each target, exhibiting a high correlation with the obtained result from a conventional real‐time PCR (qPCR) machine. Furthermore, multiplex qPCR analysis using FLG‐µSPEC and spectral deconvolution successfully demonstrates simultaneous and accurate quantification of three different targets. The FLG‐µSPEC module offers diverse on‐demand analytical applications, including not only multiplex bioassay but also environmental monitoring.

Abstract Image

用于光谱多重生物测定的平场透镜光栅微光谱仪
光谱多重生物分析法可在生物医学应用中快速、同步地检测各种生物标记物。传统光谱仪的高成本和笨重结构阻碍了用于床旁诊断的生物医学仪器的分散化。本文报告了一种基于平场透镜光栅的微光谱仪(FLG-µSPEC),用于多目标核酸扩增的连续检测。FLG-µSPEC 的特点是垂直集成微缝、集成平场透镜和透射光栅(FLG)、凹面镜和 CMOS 线传感器。FLG 通过使分散波长的焦平面平坦化,有效地校正了场曲率,同时也有利于模块的紧凑封装。FLG-µSPEC 模块可精确组装成 10 毫米 × 16 毫米 × 10 毫米的尺寸。在 500 至 950 nm 的工作范围内,平均光谱分辨率为 3.4 ± 0.9 nm。利用 FLG-µSPEC 的光谱检测系统对每个目标进行了λ DNA(𝜆-DNA)扩增,与传统实时 PCR(qPCR)设备获得的结果具有很高的相关性。此外,利用 FLG-µSPEC 和光谱解卷积技术进行的多重 qPCR 分析成功地展示了对三个不同靶标的同步精确定量。FLG-µSPEC 模块可提供多种按需分析应用,不仅包括多重生物分析,还包括环境监测。
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