Spectral phasor analysis allows rapid and reliable unmixing of fluorescence microscopy spectral images
Spectral phasor analysis allows rapid and reliable unmixing of fluorescence microscopy spectral images
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DOI：
10.1364/oe.20.012729
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发表时间：
2012-06-04
期刊：
影响因子：
3.8
通讯作者：
Gerritsen, Hans C.
中科院分区：
文献类型：
--
作者：
Fereidouni, Farzad;Bader, Arjen N.;Gerritsen, Hans C.
提出了一种新的全局分析算法来分析（超）光谱图像。它是基于相量表示，已被证明是非常强大的寿命成像数据的分析。在光谱相量分析中，图像中每个像素的荧光光谱被傅立叶变换。接下来，变换的第一谐波的真实的和虚部被用作散射（谱相量）图中的X和Y坐标。重要的是，光谱相量表示允许快速（真实的时间）半盲光谱解混的图像中的多达三个组件。这在含有三种荧光标记的固定细胞的载玻片上得到证明。此外，该方法被用于分析在新鲜的草叶片中的细胞的自发荧光。结果表明，频谱相量的方法是兼容的频谱成像数据记录与低数量的频谱通道。(c)2012美国光学学会
A new global analysis algorithm to analyse (hyper-) spectral images is presented. It is based on the phasor representation that has been demonstrated to be very powerful for the analysis of lifetime imaging data. In spectral phasor analysis the fluorescence spectrum of each pixel in the image is Fourier transformed. Next, the real and imaginary components of the first harmonic of the transform are employed as X and Y coordinates in a scatter (spectral phasor) plot. Importantly, the spectral phasor representation allows for rapid (real time) semi-blind spectral unmixing of up to three components in the image. This is demonstrated on slides with fixed cells containing three fluorescent labels. In addition the method is used to analyse autofluorescence of cells in a fresh grass blade. It is shown that the spectral phasor approach is compatible with spectral imaging data recorded with a low number of spectral channels. (c) 2012 Optical Society of America