The inherent problem of transflection-mode infrared spectroscopic microscopy and the ramifications for biomedical single point and imaging applications

被引:107
作者
Bassan, Paul [1 ,2 ]
Lee, Joe [2 ]
Sachdeva, Ashwin [1 ]
Pissardini, Juliana [1 ,2 ]
Dorling, Konrad M. [1 ,2 ]
Fletcher, John S. [1 ,2 ]
Henderson, Alex [1 ,2 ]
Gardner, Peter [1 ,2 ]
机构
[1] Univ Manchester, Manchester Inst Biotechnol, Manchester M1 7DN, Lancs, England
[2] Univ Manchester, Sch Chem Engn & Analyt Sci, Manchester M13 9PL, Lancs, England
关键词
MIE SCATTERING; BIOLOGICAL CELLS; TISSUE; MICROSPECTROSCOPY; SPECTRA; ARTIFACTS; SUPPORT; IMAGES;
D O I
10.1039/c2an36090j
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Transflection-mode FTIR spectroscopy has become a popular method of measuring spectra from biomedical and other samples due to the relative low cost of substrates compared to transmission windows, and a higher absorbance due to a double pass through the same sample approximately doubling the effective path length. In this publication we state an optical description of samples on multilayer low-e reflective substrates. Using this model we are able to explain in detail the so-called electric-field standing wave effect and rationalise the non-linear change in absorbance with sample thickness. The ramifications of this non-linear change, for imaging and classification systems, where a model is built from tissue sectioned at a particular thickness and compared with tissue of a different thickness are discussed. We show that spectra can be distorted such that classification fails leading to inaccurate tissue segmentation which may have subsequent implications for disease diagnostics applications.
引用
收藏
页码:144 / 157
页数:14
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