Search results for "Sectioning"
showing 10 items of 43 documents
Optical sectioning microscopy through single-shot Lightfield protocol
2020
Optical sectioning microscopy is usually performed by means of a scanning, multi-shot procedure in combination with non-uniform illumination. In this paper, we change the paradigm and report a method that is based in the light field concept, and that provides optical sectioning for 3D microscopy images after a single-shot capture. To do this we fi rst capture multiple orthographic perspectives of the sample by means of Fourier-domain integral microscopy (FiMic). The second stage of our protocol is the application of a novel refocusing algorithm that is able to produce optical sectioning in real time, and with no resolution worsening, in the case of sparse f luorescent samples.We provide the…
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.
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.
Optical-sectioning microscopy by patterned illumination
2010
We propose a very simple method for the flexible production of 1D structured illumination for high resolution 3D microscopy. Specifically, we propose the insertion of a Fresnel biprism after a monochromatic point source for producing a pair of twin, fully coherent, virtual point sources. The resulting interference fringes are projected into the 3D sample and, by simply varying the distance between the biprism and the point source, one can tune the period of the fringes, while keeping their contrast, in a very versatile and efficient way.
Digital slicing of 3D scenes by Fourier filtering of integral images
2008
We present a novel technique to extract depth information from 3D scenes recorded using an Integral Imaging system. The technique exploits the periodic structure of the recorded integral image to implement a Fourier-domain filtering algorithm. A proper projection of the filtered integral image permits reconstruction of different planes that constitute the 3D scene. The main feature of our method is that the Fourier-domain filtering allows the reduction of out-of-focus information, providing the InI system with real optical sectioning capacity.
Measurement and storage of a network of jacobians as a method for the visual positioning of a robot arm
1996
The goal of this paper is to describe a method to position a robot arm at any visible point of a given workspace without an explicit on line use of the analytical form of the transformations between real space and camera coordinates (camera calibration) or between cartesian and joint coordinates (direct or inverse kinematics of the robot arm). The formulation uses a discrete network of points distributed all over the workspace in which a procedure is given to measure certain Jacobian matrices which represent a good local linear approximation to the unknown compound transformation between camera and joint coordinates. This approach is inspired by the biological observation of the vestibulo-o…
Spatial calibration of structured illumination fluorescence microscopy using capillary tissue phantoms.
2008
Quantitative assessment of microvascular structure is relevant to the investigations of ischemic injury, reparative angiogenesis and tumor revascularization. In light microscopy applications, thick tissue specimens are necessary to characterize microvascular networks; however, thick tissue leads to image distortions due to out-of-focus light. Structured illumination confocal microscopy is an optical sectioning technique that improves contrast and resolution by using a grid pattern to identify the plane-of-focus within the specimen. Because structured illumination can be applied to wide-field (nonscanning) microscopes, the microcirculation can be studied by sequential intravital and confocal…
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.
Tunable optical sectioning in confocal microscopy by use of symmetrical defocusing and apodization
2008
We present two novel optical methods to achieve a significative improvement in the optical-sectioning capacity of confocal scanning microscopes. The techniques, whose real power is the simplicity with which they can be implemented, consist of a suitable combination of symmetrical defocusing with two different manners of apodizing both parts of the confocal architecture. It is shown that the proposed techniques are useful in both the bright-field and the fluorescence modes and for reflection and transmission geometries.
Optical sectioning with a Wiener-like filter in Fourier integral imaging microscopy
2018
Non-scanning, single-shot, 3D integral microscopy with optical sectioning is presented. The method is based on the combination of Fourier-mode integral microscopy with a 3D deconvolution technique. Specifically, the refocused volume provided by a regular back-projection algorithm is 3D deconvolved with a synthetic 3D impulse response function that takes into account the number and positions of the elemental images. The use of this hybrid technique provides a stack of true-color depth-refocused images with significant gain of optical sectioning. The stack can be used, among other applications, to inspect inside the thick microscope specimen, to calculate collections of perspective views wit…