Erfurth Florian, Tretyakov Alexander, Nyuyki Berla, Mrotzek Grit, Schmidt Wolf-Dieter, Fassler Dieter, Saluz Hans Peter
Department of Photonics and Sensors, Innovative Bio-, Medical- and Environmental Technologies (GMBU e.V.), Felsbachstrasse 7 D-07745 Jena, Germany.
Anal Chem. 2008 Oct 15;80(20):7706-13. doi: 10.1021/ac801014m. Epub 2008 Sep 23.
We describe the development and operation of a two-laser, large-field hyperspectral scanner for analysis of multicolor genotyping microarrays. In contrast to confocal microarray scanners, in which wavelength selectivity is obtained by positioning band-pass filters in front of a photomultiplier detector, hyperspectral microarray scanners collect the complete visible emission spectrum from the labeled microarrays. Hyperspectral scanning permits discrimination of multiple spectrally overlapping fluorescent labels with minimal use of optical filters, thus offering important advantages over standard filter-based multicolor microarray scanners. The scanner uses two-sided oblique line illumination of microarrays. Two lasers are used for the excitation of dyes in the visible and near-infrared spectral regions. The hyperspectral scanner was evaluated with commercially available two-color calibration slides and with in-house-printed four-color microarrays containing dyes with spectral properties similar to their commercial genotyping array counterparts.
我们描述了一种用于分析多色基因分型微阵列的双激光大视野高光谱扫描仪的开发和运行。与共聚焦微阵列扫描仪不同，共聚焦微阵列扫描仪通过在光电倍增管探测器前放置带通滤光片来获得波长选择性，而高光谱微阵列扫描仪则收集来自标记微阵列的完整可见发射光谱。高光谱扫描允许在最少使用光学滤光片的情况下区分多个光谱重叠的荧光标记，从而比基于标准滤光片的多色微阵列扫描仪具有重要优势。该扫描仪使用微阵列的双面斜线照明。使用两台激光器在可见光和近红外光谱区域激发染料。使用市售的双色校准载玻片和内部印刷的四色微阵列对高光谱扫描仪进行了评估，这些微阵列包含的染料的光谱特性与其商业基因分型阵列对应物相似。
