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Effects of different immersion media in multiphoton imaging of the epithelium and dermis of human skin
Author(s) -
Hsiao ChihYuan,
Sun Yen,
Chen WeiLiang,
Tung ChihKuan,
Lo Wen,
Su JiunnWen,
Lin SungJan,
Jee ShiouHwa,
Jan GwoJen,
Dong ChenYuan
Publication year - 2006
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20377
Subject(s) - dermis , oil immersion , immersion (mathematics) , epithelium , numerical aperture , refractive index , materials science , biomedical engineering , anatomy , optics , chemistry , pathology , biology , medicine , physics , optoelectronics , mathematics , composite material , wavelength , pure mathematics
In this work, we compared the performance of objectives with similar numerical aperture of 0.75 but different immersion media of air, water, glycerin, and oil in the imaging of human skin epithelium and dermis. In general, we found that the oil immersion objective recorded the strongest intensity at the same mechanical depth. We also characterized the focal shifts and found that with decreasing refractive index, the focal shift becomes increasingly more negative (for both the epithelium and dermis). In imaging the dermis, we estimated the image resolution at the depths of 18.8 and 30.2 μm, and found that the image resolution were comparable at these depths under the four types of immersion conditions. Our results demonstrate that by changing the immersion media, the main microscopic imaging effects are the recorded axial intensities and the focal shifts. The effects on the image resolution are negligible. Microsc. Res. Tech., 2006. © 2006 Wiley‐Liss, Inc.