Resonant Mie Scattering (RMieS) correction of infrared spectra from highly scattering biological samples
Resonant Mie Scattering (RMieS) correction of infrared spectra from highly scattering biological samples
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DOI：
10.1039/b921056c
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发表时间：
2010-01-01
期刊：
影响因子：
4.2
通讯作者：
Gardner, Peter
中科院分区：
文献类型：
--
作者：
Bassan, Paul;Kohler, Achim;Gardner, Peter
单个生物细胞的红外光谱通常表现出“色散伪像”，观察到吸收带的高波数侧的强度急剧下降，特别是类似于1655 cm（-1）处的酰胺I带，导致真实峰位置向下移动。这种效应的存在使得光谱的任何生物化学解释都不可靠。最近的理论揭示了“色散伪像”的起源，这归因于共振米氏散射（RMieS）。本文提出了一种修正RMieS的初步算法，并使用模拟数据进行了评估。结果表明，“色散伪影”似乎被删除，但校正是不完美的。随后实施迭代方法，由此在每次迭代之后改进参考光谱，从而导致更准确的校正。因此，校正后的光谱变得越来越能代表纯吸收光谱。使用这种校正方法可以获得可靠的峰位置。
Infrared spectra of single biological cells often exhibit the 'dispersion artefact' observed as a sharp decrease in intensity on the high wavenumber side of absorption bands, in particular the Amide I band at similar to 1655 cm(-1), causing a downward shift of the true peak position. The presence of this effect makes any biochemical interpretation of the spectra unreliable. Recent theory has shed light on the origins of the 'dispersion artefact' which has been attributed to resonant Mie scattering (RMieS). In this paper a preliminary algorithm for correcting RMieS is presented and evaluated using simulated data. Results show that the 'dispersion artefact' appears to be removed; however, the correction is not perfect. An iterative approach was subsequently implemented whereby the reference spectrum is improved after each iteration, resulting in a more accurate correction. Consequently the corrected spectra become increasingly more representative of the pure absorbance spectra. Using this correction method reliable peak positions can be obtained.