Effects of different immersion media in multiphoton imaging of the epithelium and dermis of human skin.
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 microm, 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.
Hsiao
Chih-Yuan
CY
Department of Electrical Engineering, Institute of Electro Optics, National Taiwan University, and Department of Dermatology, National Taiwan University Hospital, Taipei 106, Taiwan.
Sun
Yen
Y
Chen
Wei-Liang
WL
Tung
Chih-Kuan
CK
Lo
Wen
W
Su
Jiunn-Wen
JW
Lin
Sung-Jan
SJ
Jee
Shiou-Hwa
SH
Jan
Gwo-Jen
GJ
Dong
Chen-Yuan
CY
D020405:Dermis / Q000166:cytology*; D000078404:Epidermal Cells; D004867:Equipment Design; D006801:Humans; D021621:Imaging, Three-Dimensional; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods; D012867:Skin / Q000166:cytology*