Properties of a 4Pi confocal fluorescence microscope

• DOI: 10.1364/josaa.9.002159
• Publication Date: 2008-11-05T17:14:49Z
• Authors: Stefan Hell; Ernst H. K. Stelzer
• Keywords: 4Pi microscope; Pinhole (optics)

In a 4Pi confocal fluorescence microscope two opposing objective lenses were used to illuminate fluorescent object from both sides and collect the emissions on sides. Constructive interference of either illumination wave fronts in common focus or detection detector pinhole resulted an axial resolution approximately four times higher than that microscope. A precise uses simultaneous was built. The full width at half-maximum depth discrimination determined experimentally be 110 nm when with numerical aperture 1.4, excitation 633 nm, 725 used.