Influence of refractive-index mismatch in high-resolution three-dimensional confocal microscopy
Influence of refractive-index mismatch in high-resolution three-dimensional confocal microscopy
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DOI：
10.1364/ao.41.000685
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发表时间：
2002-02-01
期刊：
影响因子：
1.9
通讯作者：
Robello, M
中科院分区：
文献类型：
--
作者：
Diaspro, A;Federici, F;Robello, M
在共焦激光扫描显微镜的折射率失配的影响进行了广泛的研究。在荧光微球的情况下引起的轴向像差进行了测量。这些数据被用来考虑失配引起的像差，并考虑对象的大小的影响。然后重点研究了折射率失配对不同失配条件下有效系统点扩散函数和聚焦深度的影响。我们通过实验验证了点扩散函数强度分布的峰值降低，并且点扩散函数本身作为折射率失配和穿透深度的组合效应的函数逐渐变宽，导致系统的整体性能恶化。我们还通过将亚分辨率珠嵌入卵母细胞的细胞质中进行了这些相同的测量，可以认为卵母细胞的细胞质是浑浊的介质。我们发现的证据与以前开发的理论模型相一致，特别是我们发现了一个很强的依赖强度峰值的聚焦深度。(C)2002年美国光学学会。
The effects of the refractive-index mismatch in confocal laser scanning microscopy were extensively studied. The axial aberration induced in the case of fluorescent microspheres was measured. The data were used to take into account the mismatch-induced aberrations and to consider object-size influence. Then we focused on the effect of refractive-index mismatch on the effective system's point-spread function under different mismatch conditions and on depth of focusing. We experimentally verified that the peak of the point-spread function intensity profile decreases and the point-spread function itself progressively broadens as a function of the combined effect of the refractive-index mismatch and of the penetration depth, leading to a worsening of the system's overall performances. We also-performed these same measurements by embedding subresolution beads in an oocyte's cytoplasm, which can be considered a turbid medium. We found evidence consistent with the previously developed theoretical model; in particular we found a strong dependence of the intensity peak on the focusing depth. (C) 2002 Optical Society of America.