The effects of spherical aberration on multiphoton fluorescence excitation microscopy.
The effects of spherical aberration on multiphoton fluorescence excitation microscopy.
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DOI：
10.1111/j.1365-2818.2010.03449.x
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发表时间：
2011-05
影响因子：
2
通讯作者：
Dunn KW
中科院分区：
文献类型：
--
作者：
Young PA;Clendenon SG;Byars JM;Decca RS;Dunn KW
多光子荧光激发显微镜几乎总是进行样品的折射率不同的物镜浸没介质，条件，导致球面像差。由于多光子荧光激发的二次性质，预计球差将深刻影响荧光激发的深度依赖性。为了确定在多光子荧光激发显微镜的折射率失配的影响，我们测量的信号衰减，光漂白率和分辨率退化与深度在均匀的样品，最小的光散射和吸收的折射率范围内。这些研究表明，信号水平和分辨率都随着折射率不匹配样本的深度而迅速下降。光漂白率的分析表明，优势的信号衰减与深度的结果从荧光激发率降低，即使在一个系统与descandd发射收集途径。在分析嵌入大鼠肾组织中的荧光微球中获得了类似的结果，表明球差是生物样品的多光子荧光激发显微术中的重要限制因素。
Multiphoton fluorescence excitation microscopy is almost invariably conducted with samples whose refractive index differ from that of the objective immersion medium, conditions that cause spherical aberration. Due to the quadratic nature of multiphoton fluorescence excitation, spherical aberration is expected to profoundly affect the depth dependence of fluorescence excitation. In order to determine the effect of refractive index mismatch in multiphoton fluorescence excitation microscopy, we measured signal attenuation, photobleaching rates and resolution degradation with depth in homogeneous samples with minimal light scattering and absorption over a range of refractive indices. These studies demonstrate that signal levels and resolution both rapidly decline with depth into refractive index mismatched samples. Analyses of photobleaching rates indicate that the preponderance of signal attenuation with depth results from decreased rates of fluorescence excitation, even in a system with a descannd emission collection pathway. Similar results were obtained in analyses of fluorescence microspheres embedded in rat kidney tissue, demonstrating that spherical aberration is an important limiting factor in multiphoton fluorescence excitation microscopy of biological samples.
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影响因子：
3.5
作者：
Lo, W;Sun, Y;Dong, CY
通讯作者：
Dong, CY
DOI：
10.1364/josaa.13.000320
发表时间：
1996-02-01
影响因子：
1.9
作者：
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通讯作者：
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影响因子：
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通讯作者：
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影响因子：
1.9
作者：
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通讯作者：
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影响因子：
3.5
作者：
Dong, CY;Koenig, K;So, P
通讯作者：
So, P