Super-resolution scanning microscopy with virtually structured illumination

被引:0
作者
Zhang, Su [1 ]
Li, Jingtao [1 ]
Zou, Limin [1 ]
Ding, Xuemei [1 ]
机构
[1] Harbin Inst Technol, Instrument Sci & Technol, Harbin 150080, Heilongjiang, Peoples R China
来源
TENTH INTERNATIONAL SYMPOSIUM ON PRECISION ENGINEERING MEASUREMENTS AND INSTRUMENTATION | 2019年 / 11053卷
基金
中国国家自然科学基金;
关键词
Super-resolution; Structured illumination; Multiphoton processes; Scanning microscopy; Virtually structured illumination; Three-dimensional optical sectioning; LATERAL RESOLUTION; LIMIT;
D O I
10.1117/12.2511907
中图分类号
TH7 [仪器、仪表];
学科分类号
0804 ; 080401 ; 081102 ;
摘要
The resolution of optical microscopy fundamentally limited by diffraction is at best 200 nm. Super-resolution structured illumination microscopy (SR-SIM) provides an elegant way of overcoming the diffraction limit in conventional wide-field microscope by superimposing a grid pattern generated through interference of diffraction orders on the specimen while capturing images. The use of non-uniform illumination field "shift" high specimen frequencies which are out-of-band into the pass-band of the microscope through spatial frequency mixing with the illumination field. Therefore the effective bandwidth of SR-SIM is approximately twice as conventional microscopy, corresponding to a 2-fold resolution enhancement, if the difference between excitation and emission wavelength is ignored. However, such a wide-field scheme typically can only image optically thin samples and is incompatible with multiphoton processes. In this paper, we propose a Super-resolution scanning scheme with virtually structured illumination, utilizes detection sensitivity modulation on line by programming or off line by numerical processing together with temporally cumulative imaging, the excitation intensity is constant while capturing images. In this case a nondescanned array detector such as CCD camera is needed. When combined with multiphoton excitation, this scheme can image thick samples with three-dimensional optical sectioning and much improved resolution.
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页数:6
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