A new mode of contrast in biological second harmonic generation microscopy.
A new mode of contrast in biological second harmonic generation microscopy.
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DOI：
10.1038/s41598-017-13752-y
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发表时间：
2017-10-17
影响因子：
4.6
通讯作者：
Matcher SJ
中科院分区：
文献类型：
--
作者：
Green NH;Delaine-Smith RM;Askew HJ;Byers R;Reilly GC;Matcher SJ
增强的图像对比度在生物二次谐波成像显微镜（SHIM）先前已报告通过定量评估的前向epi产生的信号强度比和偏振分析。在这里，我们展示了一种新的形式的对比度：特定的材料，epi产生的二次谐波产生（SHG）的激发效率的波长依赖性，并区分胶原蛋白和肌球蛋白的比例epi产生的SHG图像在920 nm和860 nm。胶原蛋白在920 nm处显示出增加的SHG强度，而肌球蛋白在两者之间检测到的差异很小;允许SHIM表征复杂生物样品中的不同SHG生成组分。我们提出，二阶非线性结构因子的动量空间映射是这种对比的来源，并开发了一个模型的前向和epi-generated SHG波长依赖性。我们的模型表明，即使是非常小的变化，在假定的材料纤维状结构可以产生大的变化，epi产生的SHG的波长依赖性。然而，在前向SHG的情况下，尽管相同的变化影响给定波长处的绝对强度，但它们对超出预期单调下降的波长依赖性的影响非常小。我们还建议，这种差异之间的正向和epi生成的SHG提供了一个解释许多波长依赖性的差异在已发表的文献。
Enhanced image contrast in biological second harmonic imaging microscopy (SHIM) has previously been reported via quantitative assessments of forward- to epi-generated signal intensity ratio and by polarization analysis. Here we demonstrate a new form of contrast: the material-specific, wavelength-dependence of epi-generated second harmonic generation (SHG) excitation efficiency, and discriminate collagen and myosin by ratiometric epi-generated SHG images at 920 nm and 860 nm. Collagen shows increased SHG intensity at 920 nm, while little difference is detected between the two for myosin; allowing SHIM to characterize different SHG-generating components within a complex biological sample. We propose that momentum-space mapping of the second-order non-linear structure factor is the source of this contrast and develop a model for the forward and epi-generated SHG wavelength-dependence. Our model demonstrates that even very small changes in the assumed material fibrillar structure can produce large changes in the wavelength-dependency of epi-generated SHG. However, in the case of forward SHG, although the same changes impact upon absolute intensity at a given wavelength, they have very little effect on wavelength-dependency beyond the expected monotonic fall. We also propose that this difference between forward and epi-generated SHG provides an explanation for many of the wavelength-dependency discrepancies in the published literature.
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影响因子：
3.6
作者：
Hall G;Tilbury KB;Campbell KR;Eliceiri KW;Campagnola PJ
通讯作者：
Campagnola PJ
影响因子：
1.9
作者：
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通讯作者：
QUERRY, MR
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作者：
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作者：
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通讯作者：
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作者：
Deng, XY;Williams, ED;Gu, M
通讯作者：
Gu, M