Spherical aberration correction in multiphoton fluorescence imaging using objective correction collar
Spherical aberration correction in multiphoton fluorescence imaging using objective correction collar
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DOI：
10.1117/1.1924614
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发表时间：
2005-05-01
影响因子：
3.5
通讯作者：
Dong, CY
中科院分区：
文献类型：
--
作者：
Lo, W;Sun, Y;Dong, CY
多光子显微镜已经发展成为一种强大的三维生物成像工具。然而，成像生物样本的能力，深入可能会受到阻碍的样本球面像差和散射。这两种现象会导致图像分辨率下降和探测到的多光子信号丢失。在这项工作中，我们使用的校正领（盖玻片厚度）与水浸物镜，试图提高多光子成像。在我们检查的两个样品（人类皮肤和大鼠尾腱）中，我们发现，虽然图像分辨率的改善在定性上是不可见的，但测量的轴向荧光或二次谐波产生信号分布表明，使用校正环可以帮助改善检测到的多光子信号。皮肤（磺酰罗丹明B荧光）和肌腱（二次谐波产生）标本的最大增加分别为36%和57%。结果表明，对于深度多光子成像，校正环可以用来校正球差。但是，需要检查每种组织类型，以确定要使用的最佳矫正领设置。(c)2005年，由光学仪器工程师学会（Society of Photo-Optical Instrumentation Engineers）主办。
Multiphoton microscopy has evolved into a powerful bioimaging tool in three dimensions. However, the ability to image biological specimens in-depth can be hindered by sample spherical aberration and scattering. These two phenomena can result in the degradation of image resolution and the loss of detected multiphoton signal. In this work, we use the correction collar (for cover glass thickness) associated with a water immersion objective in an attempt to improve multiphoton imaging. In the two samples we examined (human skin and rat tail tendon), we found that while the improvement in image resolution was not visible qualitatively, the measured axial fluorescence or second harmonic generation signal profiles indicate that the use of the correction collar can help to improve the detected multiphoton signals. The maximum increases are 36% and 57% for the skin (sulforhodamine B fluorescence) and tendon (second harmonic generation) specimens, respectively. Our result shows that for in-depth multiphoton imaging, the correction collar may be used to correct for spherical aberration. However, each tissue type needs to be examined to determine the optimal correction collar setting to be used. (c) 2005 Society of Photo-Optical Instrumentation Engineers.