Sinclair Michael B, Timlin Jerilyn A, Haaland David M, Werner-Washburne Margaret
Sandia National Laboratories, Albuquerque, New Mexico 87185-1405, USA.
Appl Opt. 2004 Apr 1;43(10):2079-88. doi: 10.1364/ao.43.002079.
We describe the design, construction, and operation of a hyperspectral microarray scanner for functional genomic research. The hyperspectral instrument operates with spatial resolutions ranging from 3 to 30 microm and records the emission spectrum between 490 and 900 nm with a spectral resolution of 3 nm for each pixel of the microarray. This spectral information, when coupled with multivariate data analysis techniques, allows for identification and elimination of unwanted artifacts and greatly improves the accuracy of microarray experiments. Microarray results presented in this study clearly demonstrate the separation of fluorescent label emission from the spectrally overlapping emission due to the underlying glass substrate. We also demonstrate separation of the emission due to green fluorescent protein expressed by yeast cells from the spectrally overlapping autofluorescence of the yeast cells and the growth media.
我们描述了一种用于功能基因组研究的高光谱微阵列扫描仪的设计、构建和操作。该高光谱仪器的空间分辨率范围为3至30微米，可记录微阵列每个像素在490至900纳米之间的发射光谱，光谱分辨率为3纳米。当这种光谱信息与多变量数据分析技术相结合时，可用于识别和消除不需要的伪影，并大大提高微阵列实验的准确性。本研究中呈现的微阵列结果清楚地证明了荧光标记发射与由于底层玻璃基板导致的光谱重叠发射的分离。我们还展示了酵母细胞表达的绿色荧光蛋白的发射与酵母细胞和生长培养基的光谱重叠自发荧光的分离。
