Lateral spatial resolution improvement in laser scanning fluorescence microscopy using a subdiffraction limit optical spot

Takahiro Nishimura; Daiki Shinkawa; Yusuke Ogura; Yosuke Tamada; Jun Tanida

doi:10.1117/12.2319336

24 April 2018 Lateral spatial resolution improvement in laser scanning fluorescence microscopy using a subdiffraction limit optical spot

Takahiro Nishimura, Daiki Shinkawa, Yusuke Ogura, Yosuke Tamada, Jun Tanida

Author Affiliations +

Proceedings Volume 10711, Biomedical Imaging and Sensing Conference; 1071109 (2018) https://doi.org/10.1117/12.2319336
Event: SPIE Structured Light, 2018, Yokohama, Japan

Abstract

This paper presents lateral spatial resolution improvement by scanning a focused spot array pattern. To enhance the effective point spread function (PSF), we employed a phase-only computer generated hologram (CGH) that can reduce the spot size comparing to the single diffraction limited spot. A CGH was designed based on the Gerchberg-Saxton algorithm with a specific constraint to control the dispersion of light energy and the phase of generated spots. As a design example, we obtained a CGH for generating 3x3 optical spots whose sizes were reduced to 78.5% of the single diffraction limited spot. We also confirmed by simulation that the effective PSF was improved from 208 nm to 183 nm when using the subdiffraction limit spots for excitation.

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Takahiro Nishimura, Daiki Shinkawa, Yusuke Ogura, Yosuke Tamada, and Jun Tanida "Lateral spatial resolution improvement in laser scanning fluorescence microscopy using a subdiffraction limit optical spot", Proc. SPIE 10711, Biomedical Imaging and Sensing Conference, 1071109 (24 April 2018); https://doi.org/10.1117/12.2319336

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KEYWORDS

Luminescence

Diffraction

Point spread functions

Laser scanners

Spatial resolution

Computer generated holography

Microscopy

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