Fluorescence Lifetime Imaging Microscopy with Subdiffraction-Limited Resolution

被引：15
|
作者：
Lin, Po-Yen [1 ,2 ]
Lin, Yi-Cheng [1 ]
Chang, Chia-Seng [2 ]
Kao, Fu-Jen [1 ]
机构：
[1] Natl Yang Ming Univ, Inst Biophoton, Taipei 11221, Taiwan
[2] Acad Sinica, Inst Phys, Taipei 11529, Taiwan
来源：
JAPANESE JOURNAL OF APPLIED PHYSICS | 2013年 / 52卷 / 02期
关键词：
STED MICROSCOPY; STIMULATED-EMISSION; SPATIAL-RESOLUTION; NANOSCOPY; FRET;
D O I：
10.7567/JJAP.52.028004
中图分类号：
O59 [应用物理学];
学科分类号：
摘要：
In this study, we demonstrate subdiffraction-limited fluorescence lifetime imaging microscopy (FLIM) by engineering the point spread function (PSF) with stimulated emission depletion (STED). The enhanced spatial resolution allows the number of fluorophores in the PSF to reduce in turn the associated heterogeneity in lifetime analysis. Moreover, time gating can be performed using time-correlated single photon counting (TCSPC) to carefully select detected fluorescence photons so as to optimize the spatial resolution and the signal-to-noise ratio in STED imaging. This flexibility also supports the removal of the unintended effects of lifetime reduction that is caused by STED pulses. (C) 2013 The Japan Society of Applied Physics
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页数：3
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