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Journal of Biomedical Optics

Spherical aberration correction in multiphoton fluorescence imaging using objective correction collar
Author(s): Wen Lo; Yen Sun; Sung-Jan Lin; Shiou-Hwa Jee; Chen-Yuan Dong
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Paper Abstract

Multiphoton microscopy has evolved into a powerful bioimaging tool in three dimensions. However, the ability to image biological specimens in-depth can be hindered by sample spherical aberration and scattering. These two phenomena can result in the degradation of image resolution and the loss of detected multiphoton signal. In this work, we use the correction collar (for cover glass thickness) associated with a water immersion objective in an attempt to improve multiphoton imaging. In the two samples we examined (human skin and rat tail tendon), we found that while the improvement in image resolution was not visible qualitatively, the measured axial fluorescence or second harmonic generation signal profiles indicate that the use of the correction collar can help to improve the detected multiphoton signals. The maximum increases are 36% and 57% for the skin (sulforhodamine B fluorescence) and tendon (second harmonic generation) specimens, respectively. Our result shows that for in-depth multiphoton imaging, the correction collar may be used to correct for spherical aberration. However, each tissue type needs to be examined to determine the optimal correction collar setting to be used.

Paper Details

Date Published: 1 May 2005
PDF: 5 pages
J. Biomed. Opt. 10(3) 034006 doi: 10.1117/1.1924614
Published in: Journal of Biomedical Optics Volume 10, Issue 3
Show Author Affiliations
Wen Lo, National Taiwan Univ. (Taiwan)
Yen Sun, National Taiwan Univ. (Taiwan)
Sung-Jan Lin, National Taiwan Univ. Hospital (Taiwan)
Shiou-Hwa Jee, National Taiwan Univ. Hospital (Taiwan)
Chen-Yuan Dong, National Taiwan Univ. (Taiwan)

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