Sub-diffraction-limit imaging using mode multiplexing

Nan Wang; Jinping He; Jun Miyazaki; Hiromichi Tsurui; Takayoshi Kobayashi

doi:10.1117/12.2186137

21 October 2015 Sub-diffraction-limit imaging using mode multiplexing

Nan Wang, Jinping He, Jun Miyazaki, Hiromichi Tsurui, Takayoshi Kobayashi

Proceedings Volume 9636, Scanning Microscopies 2015; 96360A (2015) https://doi.org/10.1117/12.2186137
Event: SPIE Scanning Microscopies, 2015, Monterey, California, United States

Abstract

Simultaneous two-color subtraction microscopy using mode multiplexing is realized experimentally. The samples are irradiated with single laser diode at wavelength of 445 nm. Then the beam split laser spots generate separate solid and donut spatial modes and are multiplexed with modulators for simultaneous excitation. The produced fluorescence signals are back collected and further divided into two color bands with dichroic mirrors. Then they are detected with two photomultipliers and demultiplexed in four lock-in amplifiers. Four fluorescence images are recorded in every scan and resolution enhanced images are obtained in two color channels after applying the subtraction strategy. With this method, imaging results of microspheres stained with organic dyes and mesenteric lymph nodes of a mouse labeled with quantum dots (Q525/650) are realized. Improvement of 20% ~ 30% in resolving power of the two color channels compared with confocal microscopy is achieved in with corresponding subtraction factor of about 0.3.

Citation Download Citation

Nan Wang, Jinping He, Jun Miyazaki, Hiromichi Tsurui, and Takayoshi Kobayashi "Sub-diffraction-limit imaging using mode multiplexing", Proc. SPIE 9636, Scanning Microscopies 2015, 96360A (21 October 2015); https://doi.org/10.1117/12.2186137

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KEYWORDS

Luminescence

Confocal microscopy

Microscopy

Multiplexing

Image resolution

Beam splitters

Resolution enhancement technologies

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