Abstract

Optical scattering corrections are invoked to computationally distinguish between scattering and absorption contributions to recorded data in infrared (IR) microscopy, with a goal to obtain an absorption spectrum that is relatively free of the effects of sample morphology. Here, we present a modification of the extended multiplicative signal correction (EMSC) approach that allows for spectral recovery from fibers and cylindrical domains in heterogeneous samples. The developed theoretical approach is based on exact Mie theory for infinite cylinders. Although rigorous Mie theory implies utilization of comprehensive and time-consuming calculations, we propose to change the workflow of the original EMSC algorithm to minimize extensive calculations for each recorded spectrum at each iteration step. This makes the modified EMSC approach practical for routine use. First, we tested our approach using synthetic data derived from a rigorous model of scattering from cylinders in an IR microscope. Second, we applied the approach to Fourier transform IR (FT-IR) microspectroscopy data recorded from filamentous fungal and cellulose samples with pronounced fiber-like shapes. While the corrected spectra show greatly reduced baseline offsets and consistency, strongly absorbing regions of the spectrum require further refinement. The modified EMSC algorithm broadly mitigates the effects of scattering, offering a practical approach to more consistent and accurate spectra from cylindrical objects or heterogeneous samples with cylindrical domains.

© 2019 The Author(s)

Plasmonic-based impedance microspectroscopy of optically heterogeneous samples

Sidahmed A. Abayzeed
Biomed. Opt. Express 11(11) 6168-6180 (2020)

Infrared Microspectroscopy. II*

Elkan R. Blout and George R. Bird
J. Opt. Soc. Am. 41(8) 547-551 (1951)

Microspectroscopy*

John R. Loofbourow
J. Opt. Soc. Am. 40(5) 317-325 (1950)

Photon density waves scattered from cylindrical inhomogeneities: theory and experiments

Scott A. Walker, David A. Boas, and Enrico Gratton
Appl. Opt. 37(10) 1935-1944 (1998)

Correction of diffraction effects in confocal Raman microspectroscopy

A. Gallardo, R. Navarro, H. Reinecke, and S. Spells
Opt. Express 14(19) 8706-8715 (2006)