Characterizing specimen induced aberrations for high NA adaptive optical microscopy
Characterizing specimen induced aberrations for high NA adaptive optical microscopy
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DOI：
10.1364/opex.12.006540
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发表时间：
2004-12-27
期刊：
影响因子：
3.8
通讯作者：
Wilson, T
中科院分区：
文献类型：
--
作者：
Schwertner, M;Booth, MJ;Wilson, T
众所周知，像差会严重影响光学显微镜的图像质量，尤其是在共焦荧光显微镜 (CFM) 和双光子显微镜 (TPM) 中使用高数值孔径 (NA) 镜头时。自适应光学的方法可以校正像差并恢复衍射受限操作。迄今为止，生物样本成像中出现的像差问题尚未得到量化。然而，这些信息对于自适应光学系统的设计至关重要。因此，我们建造了一个包含高数值孔径物镜的干涉仪来测量生物样本引入的像差。测量的波前被分解为其泽尼克模式内容，以便对像差进行分类和量化。通过比较校正前后的信号电平，我们计算了在自适应 CFM 中使用不同 NA 校正不同数量的 Zernike 模式的潜在好处。结果表明，低阶 Zernike 模式的自适应校正可以为许多样本提供显着的好处。结果还表明，定量荧光显微镜可能会受到非自适应系统中样本引起的像差的强烈影响。 (C) 2004 年美国光学学会。
Aberrations are known to severely compromise image quality in optical microscopy, especially when high numerical aperture (NA) lenses are used in confocal fluorescence microscopy (CFM) and two-photon microscopy (TPM). The method of adaptive optics may correct aberrations and restore diffraction limited operation. So far the problem of aberrations that occur in the imaging of biological specimens has not been quantified. However, this information is essential for the design of adaptive optics systems. We have therefore built an interferometer incorporating high NA objective lenses to measure the aberrations introduced by biological specimens. The measured wavefronts were decomposed into their Zernike mode content in order both to classify and quantify the aberrations. We calculated the potential benefit of correcting different numbers of Zernike modes using different NAs in an adaptive CFM by comparing the signal levels before and after correction. The results indicate that adaptive correction of low order Zernike modes can provide significant benefit for many specimens. The results also show that quantitative fluorescence microscopy may be strongly affected by specimen induced aberrations in non-adaptive systems. (C) 2004 Optical Society of America.