Premium
Effects of objective numerical apertures on achievable imaging depths in multiphoton microscopy
Author(s) -
Tung ChihKuan,
Sun Yen,
Lo Wen,
Lin SungJan,
Jee ShiouHwa,
Dong ChenYuan
Publication year - 2004
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.20116
Subject(s) - numerical aperture , optics , microscopy , biological specimen , materials science , limiting , resolution (logic) , biomedical engineering , fluorescence lifetime imaging microscopy , fluorescence , physics , computer science , wavelength , mechanical engineering , medicine , artificial intelligence , engineering
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. Microsc. Res. Tech. 65:308–314, 2004. © 2005 Wiley‐Liss, Inc.