0000000000021853
AUTHOR
C. Ibáñez-lópez
showing 8 related works from this author
Quasi-isotropic 3-D resolution in two-photon scanning microscopy.
2009
One of the main challenges in three-dimensional microscopy is to overcome the lack of isotropy of the spatial resolution, which results from the axially-elongated shape of the point spread function. Such anisotropy gives rise to images in which significant axially-oriented structures of the sample are not resolved. In this paper we achieve an important improvement in z resolution in two-photon excitation microscopy through spatial modulation of the incident beam. Specifically, we demonstrate that the design and implementation of a simple shaded ring performs quasi-isotropic three-dimensional imaging and that the corresponding loss in luminosity can be easily compensated by most available fe…
Optical sectioning by two-pinhole confocal fluorescence microscopy.
2003
A two-pinhole axially superresolving confocal fluorescence imaging system is presented. Based on the concept of subtractive incoherent imaging, the system described here is equipped with a zero-focus complex-transmittance pupil filter in one of the collector paths. The optical sectioning capacity of the system is 25% superior to that of a free-pupil one-pinhole instrument.
Manufacture of pupil filters for 3D beam shaping
2007
In a previous work we presented a new method for binarizing pupil filters designed to control the three-dimensional (3D) irradiance distribution in the focal volume of apodized systems. The method is based in the fact that the 3D amplitude point spread function of an axially-symmetrical system can be recovered entirely from a one-dimensional (1D) set of regularly spaced amplitude samples. Hence we proposed the use of 1D iterative Fourier transform algorithm to binarize a, properly mapped, version of the amplitude transmittance of the filter. The binary masks obtained consist of a set of opaque and transparent concentric annular zones. In this paper we have built two of these masks with oppo…
Axial gain resolution in optical sectioning fluorescence microscopy by shaded-ring filters.
2009
We present a new family of pupil masks to control the axial component of the intensity distribution in the focal region of tightly focused light fields. The filters, which consist of a circular clear pupil with a single shaded ring, allow to control the width of the central lobe of the axial spot together with the residual sidelobes energy. The filters can be applied to improve the optical sectioning capacity of different scanning microscopes.
Shaded-mask filtering: novel strategy for improvement of resolution in radial-polarization scanning microscopy
2006
Radially polarized illumination produces, in combination with annular pupil filtering, the sharpest focal spot. However, the enhanced sidelobes produce an effective deterioration of spatial resolution. This study shows that if one substitutes the annular filter by one, adequately designed, shaded-mask filter, the sidelobes' strength is strongly reduced, and an important, effective improvement of resolution is achieved.
Three-ring filters increase the effective NA up to 1.46 in optical sectioning fluorescence microscopy
2003
Single-photon fluorescence confocal microscopy techniques can be combined with the use of specific binary filters in order to increase their optical sectioning capability. We present a novel class of axially super-resolving binary pupil filters specially designed to reach this aim. These filters let us to obtain a relevant compression of the z-response together with the reduction of the photo-bleaching effect typically inherent to apodization techniques. The fact of joining both the three-ring filters we propose in the illumination path, and the confocal detection gives rise to an important effective increase of lenses of effective numerical aperture.
Optical-sectioning improvement in two-color excitation scanning microscopy
2004
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.
Quasi-spherical focal spot in two-photon scanning microscopy by three-ring apodization
2005
International audience; We present a beam-shaping technique for two-photon excitation (TPE) fluorescence microscopy. We show that by inserting a properly designed three-ring pupil filter in the illumination beam of the microscope, the effective optical sectioning capacity of such a system improves so that the point spread function gets a quasi-spherical shape. Such an improvement, which allows the acquisition of 3D images with isotropic quality, is obtained at the expense of only a small increase of the overall energy in the axial sidelobes. The performance of this technique is illustrated with a scanning TPE microscopy experiment in which the image of small beads is obtained. We demonstrat…