STED with wavelengths closer to the emission maximum

被引：77

作者：

Vicidomini, Giuseppe ^{[1
]}

Moneron, Gael ^{[1
]}

Eggeling, Christian ^{[1
]}

Rittweger, Eva ^{[1
,2
]}

Hell, Stefan W. ^{[1
,2
]}

机构：

[1] Max Planck Inst Biophys Chem, Dept NanoBiophoton, D-37077 Gottingen, Germany

[2] German Canc Res Ctr BioQuant, D-69120 Heidelberg, Germany

来源：

OPTICS EXPRESS | 2012年 / 20卷 / 05期

基金：

英国医学研究理事会;

关键词：

FIELD OPTICAL NANOSCOPY; DEPLETION FLUORESCENCE MICROSCOPY; DIFFRACTION RESOLUTION LIMIT; STIMULATED-EMISSION; BREAKING; CELLS; BARRIER; DECONVOLUTION; FLUOROPHORES; GFP;

D O I：

10.1364/OE.20.005225

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

In stimulated emission depletion (STED) nanoscopy the wavelength of the STED beam is usually tuned towards the red tail of the emission maximum of the fluorophore. Shifting the STED wavelength closer to the emission peak, i.e. towards the blue region, favorably increases the stimulated emission cross-section. However, this blue-shifting also increases the probability to excite fluorophores that have remained in their ground state, compromising the image contrast. Here we present a method to exploit the higher STED efficiency of blue-shifted STED beams while maintaining the contrast in the image. The method is exemplified by imaging immunolabeled features in mammalian cells with an up to 3-fold increased STED efficiency compared to that encountered in standard STED nanoscopy implementations. (C) 2012 Optical Society of America

引用

页码：5225 / 5236

页数：12

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