한빛사 논문
Seungeun Oha,1, ChangHee Leeb,1, Wenlong Yangc, Ang Lia, Avik Mukherjeea, Markus Basana, Chongzhao Rand, Wei Yind, Clifford J. Tabinb, Dan Fue,2, X. Sunney Xief, and Marc W. Kirschnera,2
aDepartment of Systems Biology, Harvard Medical School, Boston, MA 02115; bDepartment of Genetics, Harvard Medical School, Boston, MA 02115; cCenter for Advanced Imaging, Harvard University, Cambridge, MA 20138; dAthinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02129; eDepartment of Chemistry, University of Washington, Seattle, WA 98195; and fBiomedical Pioneering Innovation Center, Peking University, Beijing 100871; China
1S.O. and C.L. contributed equally to this work.
2To whom correspondence may be addressed.
Abstract
Cell mass and chemical composition are important aggregate cellular properties that are especially relevant to physiological processes, such as growth control and tissue homeostasis. Despite their importance, it has been difficult to measure these features quantitatively at the individual cell level in intact tissue. Here, we introduce normalized Raman imaging (NoRI), a stimulated Raman scattering (SRS) microscopy method that provides the local concentrations of protein, lipid, and water from live or fixed tissue samples with high spatial resolution. Using NoRI, we demonstrate that protein, lipid, and water concentrations at the single cell are maintained in a tight range in cells under the same physiological conditions and are altered in different physiological states, such as cell cycle stages, attachment to substrates of different stiffness, or by entering senescence. In animal tissues, protein and lipid concentration varies with cell types, yet an unexpected cell-to-cell heterogeneity was found in cerebellar Purkinje cells. The protein and lipid concentration profile provides means to quantitatively compare disease-related pathology, as demonstrated using models of Alzheimer’s disease. This demonstration shows that NoRI is a broadly applicable technique for probing the biological regulation of protein mass, lipid mass, and water mass for studies of cellular and tissue growth, homeostasis, and disease.
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