Enhancement of imaging depth of two-photon microscopy using pinholes: Analytical simulation and experiments

被引：8
|
作者：
Song, Woosub [1 ]
Lee, Jihoon [2 ]
Kwon, Hyuk-Sang [1 ,2 ]
机构：
[1] Gwangju Inst Sci & Technol, Sch Informat & Mechatron, Kwangju, South Korea
[2] Gwangju Inst Sci & Technol, Dept Med Syst Engn, Kwangju 500712, South Korea
来源：
OPTICS EXPRESS | 2012年 / 20卷 / 18期
关键词：
FLUORESCENCE COLLECTION; TISSUE;
D O I：
10.1364/OE.20.020605
中图分类号：
O43 [光学];
学科分类号：
070207 ; 0803 ;
摘要：
Achieving a greater imaging depth with two-photon fluorescence microscopy (TPFM) is mainly limited by out-of-focus fluorescence generated from both ballistic and scattered light excitation. We report on an improved signal-to-noise ratio (SNR) in a highly scattering medium as demonstrated by analytical simulation and experiments for TPFM. Our technique is based on out-of-focus rejection using a confocal pinhole. We improved the SNR by introducing the pinhole in the collection beam path. Using the radiative transfer theory and the ray-optics approach, we analyzed the effects of different sizes of pinholes on the generation of the fluorescent signal in the TPFM system. The analytical simulation was evaluated by comparing its results with the experimental results in a scattering medium. In a combined confocal pinhole and two-photon microscopy system, the imaging depth limit of approximately 5 scattering mean free paths (MFP) was found to have improved to 6.2 MFP. (C) 2012 Optical Society of America
引用
收藏
页码：20605 / 20622
页数：18
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