In Situ Analysis of Living Embryonic Stem Cells by Coherent Anti-Stokes Raman Microscopy

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• 作者：Stanislav O. Konorov ; Clive H. Glover ; James M. Piret ; Jennifer Bryan ; H. Georg Schulze ; Michael W. Blades ; Robin F. B. Turner
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：September 15, 2007
• 年：2007
• 卷：79
• 期：18
• 页码：7221 - 7225
• 全文大小：168K
• 年卷期：v.79,no.18(September 15, 2007)
• ISSN：1520-6882

文摘

Embryonic stem cells (ESC), derived from preimplantation embryos, are defined by their ability to both self-renew and differentiate into all of the cells and tissues ofa mature animal. Efforts to develop methods for in vitroculture of ESC for research or eventual therapeuticapplications are hampered by the lack of rapid, nondestructive assays for distinguishing ESC from other (differentiated) cells within a growing culture. Coherent anti-Stokes Raman scattering (CARS) microscopy is shownhere to be a sensitive and nondestructive method foridentifying mouse ESC based on selective observation ofspecific molecular vibrations believed to be spectroscopicmarkers indicating the differentiated vs undifferentiatedstates of such cells. The nonlinear nature of CARS alsopermits imaging with subcellular resolution, potentiallyoffering a means by which chemical changes accompanying the early stages of differentiation may be associatedwith certain intracellular compartments (e.g., nucleus,cytoplasm, membranes). A novel exposure/collectionconfiguration is described, which yields high collectionefficiency and low interference from nonresonant background components.