Efficient fluorescence inhibition patterns for RESOLFT microscopy

被引:98
作者
Keller, Jan [1 ]
Schoenle, Andreas [1 ]
Hell, Stefan W. [1 ]
机构
[1] Max Planck Inst Biophys Chem, D-37077 Gottingen, Germany
来源
OPTICS EXPRESS | 2007年 / 15卷 / 06期
关键词
D O I
10.1364/OE.15.003361
中图分类号
O43 [光学];
学科分类号
070207 ; 0803 ;
摘要
By exploiting the saturation of a reversible single photon transition, RESOLFT microscopy is capable of resolving three dimensional structures inside specimen with a resolution that is no longer limited by the wavelength of the light in use. The transition is driven by a spatially varying intensity distribution that features at least one isolated point, line or plane with zero intensity and the resolution achieved depends critically on the field distribution around these zeros. Based on a vectorial analysis of the image formation in a RESOLFT microscope, we develop a method to effectively search for optimal zero intensity point patterns under typical experimental conditions. Using this approach, we derived a spatial intensity distribution that optimizes the focal plane resolution. Moreover, we outline a general strategy that allows optimization of the resolution for a given experimental situation and present solutions for the most common cases in biological imaging. (c) 2007 Optical Society of America.
引用
收藏
页码:3361 / 3371
页数:11
相关论文
共 25 条
[1]  
Abbe E., 1873, ARCH MIKROSK ANAT, V9, P413, DOI [10.1007/BF02956173, DOI 10.1007/BF02956173]
[2]   Generation of a hollow dark spherical spot by 4π focusing of a radially polarized Laguerre-Gaussian beam [J].
Bokor, N ;
Davidson, N .
OPTICS LETTERS, 2006, 31 (02) :149-151
[3]   2-PHOTON LASER SCANNING FLUORESCENCE MICROSCOPY [J].
DENK, W ;
STRICKLER, JH ;
WEBB, WW .
SCIENCE, 1990, 248 (4951) :73-76
[4]   Macromolecular-scale resolution in biological fluorescence microscopy [J].
Donnert, Gerald ;
Keller, Jan ;
Medda, Rebecca ;
Andrei, M. Alexandra ;
Rizzoli, Silvio O. ;
Luehrmann, Reinhard ;
Jahn, Reinhard ;
Eggeling, Christian ;
Hell, Stefan W. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (31) :11440-11445
[5]   Saturated patterned excitation microscopy - a concept for optical resolution improvement [J].
Heintzmann, R ;
Jovin, TM ;
Cremer, C .
JOURNAL OF THE OPTICAL SOCIETY OF AMERICA A-OPTICS IMAGE SCIENCE AND VISION, 2002, 19 (08) :1599-1609
[6]  
HELL S, 2006, BIOMEDICAL OPTICAL I
[7]  
HELL S, 2006, SCI MICROSCOPY
[8]   Concepts for nanoscale resolution in fluorescence microscopy [J].
Hell, SW ;
Dyba, M ;
Jakobs, S .
CURRENT OPINION IN NEUROBIOLOGY, 2004, 14 (05) :599-609
[9]   Strategy for far-field optical imaging and writing without diffraction limit [J].
Hell, SW .
PHYSICS LETTERS A, 2004, 326 (1-2) :140-145
[10]   Toward fluorescence nanoscopy [J].
Hell, SW .
NATURE BIOTECHNOLOGY, 2003, 21 (11) :1347-1355