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High efficiency beam splitter for multifocal multiphoton microscopy
Author(s) -
Nielsen T.,
Fricke M.,
Hellweg D.,
Andresen P.
Publication year - 2001
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1046/j.1365-2818.2001.00852.x
Subject(s) - optics , beam splitter , microscope , materials science , microscopy , beam (structure) , laser , microlens , optoelectronics , physics , lens (geology)
In this article we present the development of a multibeam two‐photon laser scanning microscope. A new type of beam splitter to create the multitude of laser beams is described. This type of beam splitter has higher transmission and generates more uniform beams than can be achieved with the microlens approach used by other groups. No crosstalk exists between the different foci due to small temporal delays between the individual beams. The importance of dispersion compensation to obtain maximum efficiency of the microscope is discussed. With optimum compensation the fluorescence signal was raised by a factor of 14. Different modes of detecting the fluorescence signals and their effect on imaging speed and resolution are discussed.