Abstract
We describe the three-dimensional (3-D) image formation and data acquisition in a stage scanning 4Pi confocal fluorescence microscope with the use of two-photon excitation. The 3-D point-spread functions of the 4Pi confocal and regular confocal microscope are measured and compared. Particular emphasis is given to the data acquisition procedure. 4Pi confocal microscopy results in a point-spread function that is 4 times sharper than that of a regular confocal microscope, ultimately leading to superior 3-D imaging of translucent fluorescent specimens. For a two-photon excitation wavelength of approximately 800 nm, we obtain an axial resolution of 140 nm.
© 1997 Optical Society of America
Full Article | PDF ArticleMore Like This
M. Schrader, M. Kozubek, S. W. Hell, and T. Wilson
Opt. Lett. 22(7) 436-438 (1997)
Stefan W. Hell and Matthias Nagorni
Opt. Lett. 23(20) 1567-1569 (1998)
Karsten Bahlmann and Stefan W. Hell
Appl. Opt. 39(10) 1652-1658 (2000)