Effect of a confocal pinhole in two-photon microscopy

✍ Scribed by Gauderon, R.; Lukins, P. B.; Sheppard, C. J. R.

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 300 KB
• Volume: 47
• Category: Article
• ISSN: 1059-910X
• DOI: 10.1002/(sici)1097-0029(19991101)47:3<210::aid-jemt7>3.0.co;2-h

✦ Synopsis

We investigated the effect of a finite-sized confocal pinhole on the performance of nonlinear optical microscopes based on two-photon excited fluorescence and second-harmonic generation. These techniques were implemented using a modified inverted commercial confocal microscope coupled to a femtosecond Ti:sapphire laser. Both the transverse and axial resolutions are improved when the confocal pinhole is used, albeit at the expense of the signal level. Therefore, the routine use of a confocal pinhole of optimized size is recommended for two-photon microscopy wherever the fluorescence or harmonic signals are large.