Effects of objective numerical apertures on achievable imaging depths in multiphoton microscopy.
Multiphoton microscopy is a powerful technique for achieving three-dimensional submicron imaging in biological specimens. However, specimen optical parameters such as refractive indices and scattering coefficients can result in the loss of image resolution and decreased signal in depth. These factors are coupled to the focusing objective's numerical aperture (NA) in limiting the achievable imaging depths. In this work, we performed multiphoton imaging on aqueous fluorescent solution, human skin, and rat tail tendon to show that, under the same immersion condition, lower NA objectives can examine more deeply into biological specimens and should be used when optimal imaging depths is desired.
Tung
Chih-Kuan
CK
Department of Physics, National Taiwan University, Taipei 106, Taiwan.
Sun
Yen
Y
Lo
Wen
W
Lin
Sung-Jan
SJ
Jee
Shiou-Hwa
SH
Dong
Chen-Yuan
CY
D000818:Animals; D006801:Humans; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D051381:Rats; D012867:Skin / Q000166:cytology*; D013710:Tendons / Q000166:cytology*