Research Papers

Spherical aberration correction in multiphoton fluorescence imaging using objective correction collar

[+] Author Affiliations
Wen Lo, Yen Sun

National Taiwan University, Department of Physics, Taipei 106, Taiwan

Sun-Jan Lin

National Taiwan University Hospital, Department of Dermatology, Taipei 106, Taiwan

Shiou-Hwa Jee

National Taiwan University Hospital, Department of Dermatology, Taipei 106, Taiwan

National Taiwan University College of Medicine, Department of Dermatology, Taipei 106, Taiwan

Chen-Yuan Dong

National Taiwan University, Department of Physics, Taipei 106, Taiwan

J. Biomed. Opt. 10(3), 034006 (May 26, 2005). doi:10.1117/1.1924614
History: Received Dec. 1, 2003; Revised Sep. 7, 2004; Accepted Nov. 17, 2004; May 26, 2005; Online May 26, 2005
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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. © 2005 Society of Photo-Optical Instrumentation Engineers.

© 2005 Society of Photo-Optical Instrumentation Engineers

Citation

Wen Lo ; Yen Sun ; Sun-Jan Lin ; Shiou-Hwa Jee and Chen-Yuan Dong
"Spherical aberration correction in multiphoton fluorescence imaging using objective correction collar", J. Biomed. Opt. 10(3), 034006 (May 26, 2005). ; http://dx.doi.org/10.1117/1.1924614


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