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Open AccessArticle

Quantitative Distribution of DNA, RNA, Histone and Proteins Other than Histone in Mammalian Cells, Nuclei and a Chromosome at High Resolution Observed by Scanning Transmission Soft X-Ray Microscopy (STXM)

1
School of Engineering, Tokai University, Hiratsuka, Kanagawa 259-1292, Japan
2
School of Science and Engineering, Tokyo Denki University, Hatoyama, Saitama 350-0394, Japan
3
Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai 980-8577, Japan
4
UVSOR Synchrotron, Institute Molecular Science, Okazaki, Aichi 444-8585, Japan
*
Author to whom correspondence should be addressed.
Cells 2019, 8(2), 164; https://doi.org/10.3390/cells8020164
Received: 15 December 2018 / Revised: 11 February 2019 / Accepted: 13 February 2019 / Published: 16 February 2019
(This article belongs to the Special Issue Development and Challenges in Microscopy for Cellular Imaging)
Soft X-ray microscopy was applied to study the quantitative distribution of DNA, RNA, histone, and proteins other than histone (represented by BSA) in mammalian cells, apoptotic nuclei, and a chromosome at spatial resolutions of 100 to 400 nm. The relative distribution of closely related molecules, such as DNA and RNA, was discriminated by the singular value decomposition (SVD) method using aXis2000 software. Quantities of nucleic acids and proteins were evaluated using characteristic absorption properties due to the 1s–π * transition of N=C in nucleic acids and amide in proteins, respectively, in the absorption spectra at the nitrogen K absorption edge. The results showed that DNA and histone were located in the nucleus. By contrast, RNA was clearly discriminated and found mainly in the cytoplasm. Interestingly, in a chromosome image, DNA and histone were found in the center, surrounded by RNA and proteins other than histone. The amount of DNA in the chromosome was estimated to be 0.73 pg, and the content of RNA, histone, and proteins other than histone, relative to DNA, was 0.48, 0.28, and 4.04, respectively. The method we present in this study could be a powerful approach for the quantitative molecular mapping of biological samples at high resolution. View Full-Text
Keywords: X-ray spectromicroscopy; STXM; molecular mapping; quantitative study; local content; mammalian cell; apoptotic cell; chromosome X-ray spectromicroscopy; STXM; molecular mapping; quantitative study; local content; mammalian cell; apoptotic cell; chromosome
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MDPI and ACS Style

Shinohara, K.; Toné, S.; Ejima, T.; Ohigashi, T.; Ito, A. Quantitative Distribution of DNA, RNA, Histone and Proteins Other than Histone in Mammalian Cells, Nuclei and a Chromosome at High Resolution Observed by Scanning Transmission Soft X-Ray Microscopy (STXM). Cells 2019, 8, 164. https://doi.org/10.3390/cells8020164

AMA Style

Shinohara K, Toné S, Ejima T, Ohigashi T, Ito A. Quantitative Distribution of DNA, RNA, Histone and Proteins Other than Histone in Mammalian Cells, Nuclei and a Chromosome at High Resolution Observed by Scanning Transmission Soft X-Ray Microscopy (STXM). Cells. 2019; 8(2):164. https://doi.org/10.3390/cells8020164

Chicago/Turabian Style

Shinohara, Kunio; Toné, Shigenobu; Ejima, Takeo; Ohigashi, Takuji; Ito, Atsushi. 2019. "Quantitative Distribution of DNA, RNA, Histone and Proteins Other than Histone in Mammalian Cells, Nuclei and a Chromosome at High Resolution Observed by Scanning Transmission Soft X-Ray Microscopy (STXM)" Cells 8, no. 2: 164. https://doi.org/10.3390/cells8020164

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