Effects of objective numerical apertures on achievable imaging depths in multiphoton microscopy

✍ Scribed by Chih-Kuan Tung; Yen Sun; Wen Lo; Sung-Jan Lin; Shiou-Hwa Jee; Chen-Yuan Dong

• Publisher: John Wiley and Sons
• Year: 2004
• Tongue: English
• Weight: 278 KB
• Volume: 65
• Category: Article
• ISSN: 1059-910X
• DOI: 10.1002/jemt.20116

✦ Synopsis

Abstract

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.