Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/100030
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Type: | Journal article |
Title: | High-precision pinpointing of luminescent targets in encoder-assisted scanning microscopy allowing high-speed quantitative analysis |
Author: | Zheng, X. Lu, Y. Zhao, J. Zhang, Y. Ren, W. Liu, D. Lu, J. Piper, J. Leif, R. Liu, X. Jin, D. |
Citation: | Analytical Chemistry, 2016; 88(2):1312-1319 |
Publisher: | American Chemical Society |
Issue Date: | 2016 |
ISSN: | 0003-2700 1520-6882 |
Statement of Responsibility: | Xianlin Zheng, Yiqing Lu, Jiangbo Zhao, Yuhai Zhang, Wei Ren, Deming Liu, Jie Lu, James A. Piper, Robert C. Leif, Xiaogang Liu, and Dayong Jin |
Abstract: | Compared with routine microscopy imaging of a few analytes at a time, rapid scanning through the whole sample area of a microscope slide to locate every single target object offers many advantages in terms of simplicity, speed, throughput, and potential for robust quantitative analysis. Existing techniques that accommodate solid-phase samples incorporating individual micrometer-sized targets generally rely on digital microscopy and image analysis, with intrinsically low throughput and reliability. Here, we report an advanced on-the-fly stage scanning method to achieve high-precision target location across the whole slide. By integrating X- and Y-axis linear encoders to a motorized stage as the virtual "grids" that provide real-time positional references, we demonstrate an orthogonal scanning automated microscopy (OSAM) technique which can search a coverslip area of 50 × 24 mm(2) in just 5.3 min and locate individual 15 μm lanthanide luminescent microspheres with standard deviations of 1.38 and 1.75 μm in X and Y directions. Alongside implementation of an autofocus unit that compensates the tilt of a slide in the Z-axis in real time, we increase the luminescence detection efficiency by 35% with an improved coefficient of variation. We demonstrate the capability of advanced OSAM for robust quantification of luminescence intensities and lifetimes for a variety of micrometer-scale luminescent targets, specifically single down-shifting and upconversion microspheres, crystalline microplates, and color-barcoded microrods, as well as quantitative suspension array assays of biotinylated-DNA functionalized upconversion nanoparticles. |
Rights: | © 2015 American Chemical Society |
DOI: | 10.1021/acs.analchem.5b03767 |
Grant ID: | http://purl.org/au-research/grants/arc/LE150100177 http://purl.org/au-research/grants/arc/LP140100462 http://purl.org/au-research/grants/arc/FT130100517 |
Published version: | http://dx.doi.org/10.1021/acs.analchem.5b03767 |
Appears in Collections: | Aurora harvest 3 IPAS publications |
Files in This Item:
File | Description | Size | Format | |
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hdl_100030.pdf | Submitted version | 1.01 MB | Adobe PDF | View/Open |
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