Imaging properties of high aperture multiphoton fluorescence scanning optical microscopes
Imaging properties of high aperture multiphoton fluorescence scanning optical microscopes
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DOI：
10.1046/j.1365-2818.1999.00448.x
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发表时间：
1999-02
影响因子：
2
通讯作者：
P. Higdon;Peter Török;Tony Wilson
中科院分区：
文献类型：
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作者：
P. Higdon;Peter Török;Tony Wilson
本文提出了一种采用有限尺寸探测器的大孔径扫描光学显微镜多光子荧光成像理论。这种显微镜的成像性能的荧光发射的偏振的效果进行了研究。计算了对四联苯单光子、双光子和三光子激发下高、低孔径光学系统的横向和轴向分辨率。横向分辨率的显着改善被发现通过采用共焦针孔实现。这种改进随着多光子过程的阶数而增加。同时，发现当针孔的尺寸减小以实现最佳分辨率时，信噪比不会降低超过30%。发现使用共焦检测可以显著提高光学切片的程度。对于双光子和三光子激发，预测轴向全宽半峰提高30%。
A theory for multiphoton fluorescence imaging in high aperture scanning optical microscopes employing finite sized detectors is presented. The effect of polarisation of the fluorescent emission on the imaging properties of such microscopes is investigated. The lateral and axial resolutions are calculated for one‐, two‐ and three‐photon excitation of p‐quaterphenyl for high and low aperture optical systems. Significant improvement in lateral resolution is found to be achieved by employing a confocal pinhole. This improvement increases with the order of the multiphoton process. Simultaneously, it is found that, when the size of the pinhole is reduced to achieve the best possible resolution, the signal‐to‐noise ratio is not degraded by more than 30%. The degree of optical sectioning achieved is found to improve dramatically with the use of confocal detection. For two‐ and three‐photon excitation axial full width half‐maximum improvement of 30% is predicted.