FTIR microscopy of biological cells and tissue: data analysis using resonant Mie scattering (RMieS) EMSC algorithm

Area of Science:

• Spectroscopy
• Biomedical Imaging
• Computational Chemistry

Background:

• Infrared microscopy of biological samples is hindered by scattering, specifically resonant Mie scattering (RMieS).
• Scattering effects distort spectral baselines, peak shapes, and positions, complicating data interpretation.
• Accurate analysis of biological tissues using infrared microscopy requires effective mitigation of these scattering artifacts.

Purpose of the Study:

• To introduce and evaluate the resonant Mie scattering extended multiplicative signal correction (RMieS-EMSC) algorithm.
• To assess the impact of user-defined parameters (iterations, reference spectrum) on RMieS-EMSC accuracy and computational time.
• To demonstrate the algorithm's utility in analyzing infrared images of biological tissues.

Main Methods:

• Development of the RMieS-EMSC algorithm for baseline and scattering correction.
• Systematic evaluation of the algorithm's performance with varying numbers of iterations.
• Investigation of the influence of different initial reference spectra on the correction outcome.
• Application of the algorithm to infrared microscopy data from biological tissues.

Main Results:

• RMieS-EMSC effectively removes scattering distortions in infrared microscopy spectra.
• Image classification of tissues is achievable with fewer than 10 iterations.
• Accurate biochemical interpretation may necessitate more iterations.
• The choice of a non-ideal reference spectrum does not prevent obtaining a unique correction solution.
• Similar reference spectra reduce the number of iterations required for accurate correction.

Conclusions:

• The RMieS-EMSC algorithm provides a robust solution for scattering artifacts in infrared microscopy.
• Parameter selection influences computational efficiency and interpretive accuracy.
• The algorithm facilitates reliable biochemical analysis of biological tissues from infrared imaging data.