We report the design and implementation of a laser scanning confocal fluorescence system with spectroscopy and anisotropy imaging capabilities. Confocal spectroscopy is achieved with a fiber pinhole that is inserted into and removed from the detection path as needed. Fluorescence anisotropy imaging is accomplished with a polarizing beam splitter placed after the conventional pinhole. Two orthogonal polarizations are detected simultaneously with balanced photomultiplier tubes. The quality of the axial sectioning that is achieved in the confocal fluorescence spectroscopy mode is demonstrated experimentally, and examples of polarization-sensitive fluorescence imaging are demonstrated in tumor cell monolayers.
© 2003 Optical Society of America
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.1790) Medical optics and biotechnology : Confocal microscopy
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.5180) Medical optics and biotechnology : Photodynamic therapy
(170.6280) Medical optics and biotechnology : Spectroscopy, fluorescence and luminescence
Chad E. Bigelow, David L. Conover, and Thomas H. Foster, "Confocal fluorescence spectroscopy and anisotropy imaging system," Opt. Lett. 28, 695-697 (2003)