Reduction of the spherical aberration effect in high-numerical-aperture optical scanning instruments.
In modern high-numerical-aperture (NA) optical scanning instruments, such as scanning microscopes, optical data storage systems, or laser trapping technology, the beam emerging from the high-NA objective focuses deeply through an interface between two media of different refractive index. Such a refractive index mismatch introduces an important amount of spherical aberration, which increases dynamically when scanning at increasing depths. This effect strongly degrades the instrument performance. Although in the past few years many different techniques have been reported to reduce the spherical aberration effect, no optimum solution has been found. Here we concentrate on a technique whose mai…
New Analytical Tools for Evaluation of Spherical Aberration in Optical Microscopy
The required tightly focused spots in three-dimensional (3D) scanning optical techniques are usually achieved by high-NA immersion lenses. The refractive index mismatch between the sample and the immersion medium introduces an important amount of spherical aberration when imaging deep inside the specimen, spreading out the focusing response. Since this aberration depends on the focalization depth, it is not possible to simultaneously achieve a global compensation for the whole scanned sample. In this way, the design of pupil elements that increase the tolerance to this aberration is of great interest. We present a new formalism for the evaluation and the design of filters that decrease the …
Shaded-Mask Filtering for Extended Depth-of-Field Microscopy
This paper proposes a new spatial filtering approach for increasing the depth-of-field (DOF) of imaging systems, which is very useful for obtaining sharp images for a wide range of axial positions of the object. Many different techniques have been reported to increase the depth of field. However the main advantage in our method is its simplicity, since we propose the use of purely absorbing beam-shaping elements, which allows a high focal depth with a minimum modification of the optical architecture. In the filter design, we have used the analogy between the axial behavior of a system with spherical aberration and the transverse impulse response of a 1D defocused system. This allowed us the…
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.
Reduction of focus size in tightly focused linearly polarized beams
The electromagnetic theory predicts that when a linearly polarized collimated field is focused by a high-angle focusing system, components perpendicular to the initial polarization are generated. The use of annular masks to reduce the area of the focal spot usually increases the magnitude of this phenomenon, known as depolarization. We present a class of masks, the three-ring masks, which are important because they narrow the central lobe of the focal intensity distribution without increasing the depolarization. This can be very useful in modern optical applications, such as confocal microscopy or multiphoton scanning microscopy.
Simple demonstration of the impact of spherical aberration on optical imaging
We present an experiment, well adapted for students of introductory optics courses, for the visualization of the impact of spherical aberration in the point spread function of imaging systems. The demonstrations are based on the analogy between the point-spread function of spherically aberrated systems, and the defocused patterns of 1D slit-like screens.