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Originally published as Biophys J. BioFAST on March 4, 2005.
doi:10.1529/biophysj.104.057950
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Biophysical Journal 88:3635-3640 (2005)
© 2005 The Biophysical Society

Mie-Type Scattering and Non-Beer-Lambert Absorption Behavior of Human Cells in Infrared Microspectroscopy

Brian Mohlenhoff *, Melissa Romeo *, Max Diem * and Bayden R. Wood {dagger}

* Department of Chemistry and Biochemistry, City University of New York, Hunter College, New York, New York; and {dagger} Centre for Biospectroscopy and School of Chemistry, Monash University, Clayton, Victoria, Australia

Correspondence: Address reprint requests to Max Diem, Hunter College, CUNY, Dept. of Chemistry, 695 Park Ave., New York, NY 10021. Tel.: 212-772-5359; E-mail: mdiem{at}hunter.cuny.edu.

We report infrared microspectral features of nuclei in a completely inactive and contracted (pyknotic) state, and of nuclei of actively dividing cells. For pyknotic nuclei, the very high local concentration of DNA leads to opaqueness of the chromatin and, consequently, the absence of DNA signals in the IR spectra of very small nuclei. However, these nuclei can be detected by their scattering properties, which can be described by the Mie theory of scattering from dielectric spheres. This scattering depends on the size of the nucleus; consequently, quite different scattering cross-sections are calculated and observed for pyknotic and mitotic nuclei.




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