The point-spread function (PSF) plays a fundamental role in deciding the resolution of an optical microscope. One way to find the experimental PSF is to image sub-resolution nanobead. However, the PSF can also be approximated numerically by knowing the optical arrangement of the imaging system. In this paper, we estimate the experimental three-dimensional PSF of a widefield microscope by taking several z-stacks of an arbitrary target, followed by application of our 2D PSF estimation scheme to each z slice. Here we use a standard resolution test target as the arbitrary target whose geometrical-optics predicted image can be easily constructed.
Zernike polynomials are orthogonal polynomials that form a complete basis set and can be easily used to describe aberrations present in an optical system. Zernike modes find applications in various fields like adaptive optics (AO), optical imaging, ophthalmology, free space optical (FSO) communication, etc. Since the modes are orthogonal, they can express any arbitrary wavefront as their linear combinations. The orthogonality of the modes enables the calculation of the expansion coefficients and suggests the independent behaviour of the Zernike mode. In this work, we numerically estimate the wavefront, defined as Zernike modes, using various state of the art phase retrieval methods. We use the Zonal wavefront sensor (ZWFS) and Transport of Intensity Equation (TIE) for phase reconstruction and then calculate the orthogonality between reconstructed Zernike modes. It is found that the reconstructed Zernike modes are not perfectly orthogonal, which is mainly due to the discrete representation of the Zernike modes. We further investigate how the change in the number of zones in a ZWFS affects orthogonality. We also simulate TIE to retrieve the phase and compare the orthogonality results with ZWFS. This study will be helpful in applications where a wavefront described using Zernike mode needs to be reconstructed, and improvement in the orthogonality is required, which is achieved by increasing the number of zones in the ZWFS and representing Zernike modes in a more continuous form.
The wavefront measurement accuracy of a grating array based zonal wavefront sensor (GAWS) can be affected by the non-uniform focal spot array and unwanted orders in the detector plane. The non-uniform focal spot array is the outcome of the non-uniform nature of the incident illumination beam’s intensity profile. This paper describes a method that dynamically modulates the laser beam’s intensity using computer generated holography, making the focal spot array uniform and eliminating unwanted spots in a detector plane, thereby enhancing the accuracy of the wavefront measurement. Here, we present proof-of-principle simulation results that demonstrate the working of the proposed improvements in GAWS.
Astigmatism has a very unique ability to change the shape of the intensity profile of an optical beam as we move away on either side of the focal plane of the optical system. This property of astigmatism can be used to measure the focussing error or the amplitude of defocus present at the plane of measurement. The use of astigmatism to measure focussing error is a very simple and easily implementable process. Astigmatism can be introduced with the help of an astigmatic lens. The approach provides a direct measure of the amplitude of defocus present in an optical beam from the measure of the intensity at the focal plane. In this paper we put forward a theoretical discussion on the above mentioned approach.
One important parameter in the case of translation of an optically trapped particle is the maximum achievable speed of translation in optical trapping. While the parameter is expected to have a dependence on the particle diameter and the viscosity of the medium, there will also be dependence on the laser power and step size of the moving trap. In this paper, we will experimentally investigate the maximum translation speed of a given trapped particle in a certain medium achievable in a holographic optical trap. We will implement the holographic trap using a liquid crystal spatial light modulator with a computer interface and use latex beads in water for trapping.
The incorporation of polarization control on the illumination beam in a laser scanning confocal microscope allows extraction of the orientational information of submicroscopic features of a sample being studied. In this paper, we present the implementation of an optical arrangement that generates homogeneous as well as non-homogeneous user define polarization profiles over the cross-sectional area of a laser beam. A confocal system is built with this optical arrangement to obtain images of the same target with different polarized illumination beams such that there is considerable reduction in the time gap between two consecutive illuminations of each location of the sample.
The high resolution applications of a laser scanning imaging system very much demand the accurate positioning of the illumination beam. The galvanometer scanner based beam scanning imaging systems, on the other hand, suffer from both short term and long term beam instability issues. Fortunately Computer generated holography based beam scanning offers extremely accurate beam steering, which can be very useful for imaging in high-resolution applications in confocal microscopy. The holographic beam scanning can be achieved by writing a sequence of holograms onto a spatial light modulator and utilizing one of the diffracted orders as the illumination beam. This paper highlights relative advantages of such a holographic beam scanning based confocal system and presents some of preliminary experimental results.
The estimation of the Point Spread Function (PSF) of an imaging system is important for various post acquisition processes. The PSF can be estimated by knowing the optical arrangement of the imaging system or can be obtained by using a point object. Both the techniques have their own limitations. In this paper we propose a new PSF estimation technique based on a target that can be reconfigured programmably. We will show that a target with different illumination areas can be imaged to establish a relation between the image plane and the object plane via a PSF. The relation thus allows one to estimate the PSF of the imaging system.
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