Optical-sectioning improvement in two-color excitation scanning microscopy.
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.
Ibáñez-López
Cristina
C
Department of Optics, University of Valencia, E46100 Burjassot, Spain.
Escobar
Isabel
I
Saavedra
Genaro
G
Martínez-Corral
Manuel
M
D004867:Equipment Design; D005374:Filtration / Q000295:instrumentation*; D007089:Image Enhancement / Q000295:instrumentation; D021621:Imaging, Three-Dimensional / Q000295:instrumentation / Q000379:methods; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation* / Q000379:methods