Infrared ATR: a probe for cellular activation

Jerilyn A. Timlin; Laura E. Martin; M. Kathleen Alam; David M. Haaland; Kristen Garrison; C. Richard Lyons; Brian Hjelle

doi:10.1117/12.455749

22 February 2002 Infrared ATR: a probe for cellular activation

Jerilyn A. Timlin, Laura E. Martin, M. Kathleen Alam, David M. Haaland, Kristen Garrison, C. Richard Lyons, Brian Hjelle

Author Affiliations +

Proceedings Volume 4577, Vibrational Spectroscopy-based Sensor Systems; (2002) https://doi.org/10.1117/12.455749
Event: Environmental and Industrial Sensing, 2001, Boston, MA, United States

Abstract

We employ infrared spectroscopy (IR) with attenuated total reflectance (ATR) as a sampling technique to monitor live and dried RAW cells (a murine macrophage cell line) during activation with g-interferon and lipopolysaccharide. By comparing the spectra of activated cells at various time points to the spectra of healthy control cells, we identify spectral bands associated with nucleic acids that are markers for the cell activation process. These spectral changes are slight and can be complicated with the normal metabolic changes that occur within cells. We will discuss the use of data pretreatment strategies to accurately correct for the contribution of the buffer to the live cell spectra. We find the standard background correction method inadequate for concentrated solutions of cells. Data presented shows the severe effect incorrect background subtraction has on the cell spectra. We report a more accurate correction for phosphate buffer spectral contribution using an interactive subtraction of the buffer spectrum. We will show classification of dried control and activated macrophage cell spectra using partial-least squares analysis with multiplicative scatter correction.

Citation Download Citation

Jerilyn A. Timlin, Laura E. Martin, M. Kathleen Alam, David M. Haaland, Kristen Garrison, C. Richard Lyons, and Brian Hjelle "Infrared ATR: a probe for cellular activation", Proc. SPIE 4577, Vibrational Spectroscopy-based Sensor Systems, (22 February 2002); https://doi.org/10.1117/12.455749

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KEYWORDS

Crystals

Infrared spectroscopy

Attenuated total reflectance

Proteins

Infrared radiation

Absorbance

FT-IR spectroscopy

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