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Optical‐sectioning improvement in two‐color excitation scanning microscopy
Author(s) -
IbáñezLópez Cristina,
Escobar Isabel,
Saavedra Genaro,
MartínezCorral Manuel
Publication year - 2004
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20073
Subject(s) - optical sectioning , microscopy , optical microscope , optics , materials science , biophysics , chemistry , scanning electron microscope , biology , physics
We present a new beam‐shaping technique for two‐color excitation fluorescence microscopy. We show that by simply inserting a properly designed shaded‐ring filter in the illumination beam of smaller wavelength, it is possible to improve the effective optical sectioning capacity of such microscopes by 23%. Such an improvement is obtained at the expense of only a very small increasing of the overall energy in the point‐spread‐function sidelobes. The performance of this technique is illustrated by a numerical imaging simulation. Microsc. Res. Tech. 64:96–102, 2004. © 2004 Wiley‐Liss, Inc.