Quantitative characterization of biological liquids for third-harmonic generation microscopy
Quantitative characterization of biological liquids for third-harmonic generation microscopy
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DOI：
10.1529/biophysj.106.094946
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发表时间：
2007-01-01
影响因子：
3.4
通讯作者：
Beaurepaire, Emmanuel
中科院分区：
文献类型：
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作者：
Debarre, Delphine;Beaurepaire, Emmanuel
三次谐波生成（THG）显微镜提供了基于三阶非线性磁化率、折射率和色散的空间变化的未染色生物样品的图像。在本研究中，我们建立了几种溶剂（水、乙醇、甘油）、生理水（离子、氨基酸、多肽、牛血清白蛋白、葡萄糖）和脂质（甘油三酯、胆固醇）溶液的三阶非线性敏感性的定量值，作为溶质浓度在1.05-1.25 μ m激发范围内的函数。我们将这些数据与成像实验相结合，表明类似1.2 mm激发的THG成像在检测生理离子浓度变化方面缺乏特异性和敏感性，而非水结构（如脂质体）提供了更强大的信号来源。最后，我们说明了索引匹配液体在THG图像中的影响。这些数据为解释生物THG图像和开发其他应用提供了基础。
Third-harmonic generation (THG) microscopy provides images of unstained biological samples based on spatial variations in third-order nonlinear susceptibility, refractive index, and dispersion. In this study, we establish quantitative values for the third-order nonlinear susceptibilities of several solvents (water, ethanol, glycerol), physiological aqueous (ions, amino acids, polypeptides, bovine serum albumin, glucose) and lipid (triglycerides, cholesterol) solutions as a function of solute concentration in the 1.05-1.25 mu m excitation range. We use these data in conjunction with imaging experiments to show that THG imaging with similar to 1.2 mm excitation lacks specificity and sensitivity to detect physiological ion concentration changes, and that nonaqueous structures such as lipid bodies provide a more robust source of signal. Finally, we illustrate the impact of index-matching liquids in THG images. These data provide a basis for interpreting biological THG images and for developing additional applications.