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. 2019 Jul 25;47(18):e108. doi: 10.1093/nar/gkz630

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© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — The AFM-IR results achieved for the typical human chromosome isolated from HeLa cell: (A) AFM topography, (B) the contact resonance frequency of AFM cantilever correlated to the local stiffness to demonstrate that the banding contrast is related with chemical structure not local stiffness heterogeneity, (C) the integrated area of an absorption band at 1240 cm⁻¹ corresponding to the O-P-O asymmetric stretching vibration, (D) the integrated area of an absorption band at 2952 cm⁻¹ corresponding to the vibration of methyl (-CH₃) asymmetric stretching vibration, (E) the ratio of the integrated absorption band at 2952 cm⁻¹ to the absorption band at 1240 cm⁻¹, (F) anty 5-methylcytosine direct immunostaining counterstained with propidium iodide, similarities between (E) and (F) can be observed: high intensity of signal from 5- methylcytosine (F) correspond to high CH₃/OPO ratio (E, yellow), (G) a direct comparison of profiles extracted along red (AFM-IR) and blue (fluorescence) lines; chromosome thickness: 190 – 210 nm, pixel size 7.8 × 7.8 nm.