Silicon nitride as a versatile growth substrate for microspectroscopic imaging and mapping of individual cells
Silicon nitride as a versatile growth substrate for microspectroscopic imaging and mapping of individual cells
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DOI：
10.1039/c001499k
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发表时间：
2010-01-01
影响因子：
--
通讯作者：
Lay, Peter A.
中科院分区：
文献类型：
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作者：
Carter, Elizabeth A.;Rayner, Benjamin S.;Lay, Peter A.
本文描述了一种通用的采样方案，该方案包括培养、分化和。在一个样品衬底(Si(3)N(4))上Xing细胞的首选形态，以使随后的各种现代显微光谱分析成为可能。该方案提供了关于代谢过程、疾病及其治疗的细胞内生物化学的前所未有的相关和补充信息，这为在分子水平上彻底改变我们对细胞和组织生物学的理解提供了机会。将贴壁细胞培养到廉价的Si3N4膜上，可以进行从硬X射线荧光(XRF和XANES)到可见光和荧光显微镜以及红外显微光谱分析，而不受基质干扰。贴壁哺乳动物细胞系(3T3-L1脂肪细胞和H9c2心肌细胞)说明了这些方案的体外应用。细胞强烈附着在Si(3)N(4)膜上，可见正常的增殖和表型生长；更重要的是，细胞的快速酒精固定不影响其结构完整性，用于后续分析。
Herein is described a general sampling protocol that includes culture, differentiation and. xing of cells in their preferred morphology on the one sample substrate (Si(3)N(4)) to enable subsequent diverse modern microspectroscopic analyses. The protocol enables unprecedented correlated and complementary information on the intracellular biochemistry of metabolic processes, diseases and their treatment, which offers the opportunity to revolutionize our understanding of cell and tissue biology at a molecular level. The culture of adherent cells onto inexpensive Si3N4 membranes allows microspectroscopic analyses across the electromagnetic spectrum, from hard X-ray fluorescence (both XRF and XANES), through to visible and fluorescence light microscopies, and infrared microspectroscopy without substrate interference. Adherent mammalian cell lines (3T3-L1 adipocytes and H9c2 cardiac myocytes) illustrate the in vitro application of these protocols. The cells adhered strongly to Si(3)N(4) membranes and visually displayed normal proliferative and phenotypic growth; more importantly, rapid alcohol fixation of cells did not affect their structural integrity for subsequent analyses.