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Journal of Biomedical Optics • Open Access

Saturated excitation microscopy for sub-diffraction-limited imaging of cell clusters

Paper Abstract

Saturated excitation (SAX) microscopy offers high-depth discrimination predominantly due to nonlinearity in the fluorescence response induced by the SAX. Calculation of the optical transfer functions and the edge responses for SAX microscopy revealed the contrast improvement of high-spatial frequency components in the sample structure and the effective reduction of background signals from the out-of-focus planes. Experimental observations of the edge response and x-z cross-sectional images of stained HeLa cells agreed well with theoretical investigations. We applied SAX microscopy to the imaging of three-dimensional cultured cell clusters and confirmed the resolution improvement at a depth of 40 μm. This study shows the potential of SAX microscopy for super-resolution imaging of deep parts of biological specimens.

Paper Details

Date Published: 2 December 2013
PDF: 8 pages
J. Biomed. Opt. 18(12) 126002 doi: 10.1117/1.JBO.18.12.126002
Published in: Journal of Biomedical Optics Volume 18, Issue 12
Show Author Affiliations
Masahito Yamanaka, Osaka Univ. (Japan)
Yasuo Yonemaru, Osaka Univ. (Japan)
Shogo Kawano, Osaka Univ. (Japan)
Kumiko Uegaki, Osaka Univ. (Japan)
Nicholas I. Smith, Osaka Univ. (Japan)
Satoshi Kawata, Osaka Univ. (Japan)
Katsumasa Fujita, Osaka Univ. (Japan)

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