Protein and lipid mass concentration measurement in tissues by stimulated Raman scattering microscopy.
Protein and lipid mass concentration measurement in tissues by stimulated Raman scattering microscopy.
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用受激拉曼散射显微镜测量组织中蛋白质和脂肪的质量浓度。
DOI：
10.1073/pnas.2117938119
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发表时间：
2022-04-26
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
--
作者：
我们报告了一种定量拉曼显微镜方法，该方法以高空间分辨率测量活体和固定组织样本中细胞中蛋白质和脂质的浓度，从而可以定量研究细胞培养物和组织切片中的细胞大小和细胞器调节；它可以应用于细胞生长、细胞分化、细胞衰老和病理学背景下的细胞大小控制、细胞内拥挤和脂质代谢问题。细胞质量和化学成分是重要的聚集细胞特性，与生长控制和组织稳态等生理过程特别相关。尽管它们很重要，但很难在完整组织的单个细胞水平上定量测量这些特征。在这里，我们介绍标准化拉曼成像 (NoRI)，这是一种受激拉曼散射 (SRS) 显微镜方法，可提供活体或固定组织样本中蛋白质、脂质和水的局部浓度，具有高空间分辨率。使用NoRI，我们证明，在相同的生理条件下，单细胞中的蛋白质、脂质和水浓度在细胞中维持在严格的范围内，并且在不同的生理状态下发生改变，例如细胞周期阶段、附着于不同硬度的基质或进入衰老。在动物组织中，蛋白质和脂质浓度随细胞类型的不同而变化，但在小脑浦肯野细胞中发现了意想不到的细胞间异质性。蛋白质和脂质浓度分布提供了定量比较疾病相关病理学的方法，正如阿尔茨海默病模型所证明的那样。该演示表明，NoRI 是一种广泛适用的技术，可用于探索蛋白质质量、脂质质量和水质量的生物调节，以研究细胞和组织生长、稳态和疾病。
We report a quantitative Raman microscopy method that measures the concentration of protein and lipid in cells at high spatial resolution in living and in fixed samples of tissues, allowing quantitative studies of cell size and organelle regulation both in cell culture and in tissue slices; it can be applied to problems of cell size control, intracellular crowding, and lipid metabolism in the context of cell growth, cell differentiation, cell senescence, and pathology. 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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影响因子：
48
作者：
Godin, Michel;Delgado, Francisco Feijo;Son, Sungmin;Grover, William H.;Bryan, Andrea K.;Tzur, Amit;Jorgensen, Paul;Payer, Kris;Grossman, Alan D.;Kirschner, Marc W.;Manalis, Scott R.
通讯作者：
Manalis, Scott R.
影响因子：
3.3
作者：
Perez Gonzalez N;Tao J;Rochman ND;Vig D;Chiu E;Wirtz D;Sun SX
通讯作者：
Sun SX
影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1126/science.1245075
发表时间：
2015-05-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Ginzberg MB;Kafri R;Kirschner M
通讯作者：
Kirschner M
DOI：
10.1073/pnas.1114477108
发表时间：
2011-11-01
影响因子：
11.1
作者：
Dill, Ken A.;Ghosh, Kingshuk;Schmit, Jeremy D.
通讯作者：
Schmit, Jeremy D.