High-Speed Lens-Free Holographic Sensing of Protein Molecules Using Quantitative Agglutination Assays.
High-Speed Lens-Free Holographic Sensing of Protein Molecules Using Quantitative Agglutination Assays.
复制标题
使用定量凝集测定法对蛋白质分子的无透镜全息感应。
DOI：
10.1021/acssensors.0c02481
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发表时间：
2021-03-26
期刊：
影响因子：
8.9
通讯作者：
McLeod E
中科院分区：
文献类型：
--
作者：
Xiong Z;Potter CJ;McLeod E
Accurate, cost-effective, easy-to-use, and point-of-care sensors for protein biomarker levels are important for disease diagnostics. A cost-effective and compact readout approach that has been used for several diagnostic applications is lens-free holographic microscopy, which provides an ultra-large field of view and submicron resolution when it is coupled with pixel superresolution techniques. Despite its potential as a diagnostic technique, lens-free microscopy has not previously been applied to quantitative protein molecule sensing in solution, which can simplify sensing protocols and ultimately enable measurements of binding kinetics in physiological conditions. Here we sense interferon-gamma (an immune system biomarker) and NeutrAvidin molecules in solution by combining lens-free microscopy with a one-step bead-based agglutination assay, enabled by a custom high-speed LED array and automated image processing routines. We call this a quantitative large-area binding (QLAB) sensor. The high-speed light source provides, for the first time, pixel-superresolved imaging of > 104 2-micron beads in solution undergoing Brownian motion, without significant motion blur. The automated image processing routines enable the counting of individual beads and clusters, providing a quantitative sensor readout that depends on both bead and analyte concentrations. Fits to chemical binding theory are provided. For NeutrAvidin, we find a limit of detection (LOD) of < 27 ng/mL (450 pM) and a dynamic range of 2–4 orders of magnitude. For mouse interferon-gamma, the LOD is < 3 ng/mL (200 pM) and the dynamic range is at least 4 orders of magnitude. The QLAB sensor holds promise for point-of-care applications in low-resource communities and where protocol simplicity is important.
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