Dual-channel phase-shifting interferometry for microscopy with second wavelength assistance

Dual-channel phase-shifting interferometry for simultaneous phase microscopy is presented. Red and blue light beams are used for microscope illumination. A 45 tilted beamsplitter replicates the object and reference waves in red light together with the object wave in blue light into two parallel beams. The two resulting quadrature phase-shifting interferograms in red light and the object waves in blue light are generated in the two channels. The two interferograms are recorded simultaneously by a color charge-coupled device (CCD) camera, and can be separated via RGB components of the recorded color patterns without crosstalk. As a result, the phase of tested specimen can be retrieved. The feasibility of the proposed method is demonstrated by test performed on a microscopic specimen.     

Physical spherical phase compensation in reflection digital holographic microscopy

Reflection configured digital holographic microscopy (DHM) can perform accurate optical topography measurements of reflecting objects, such as MEMs, MOEMs, and semiconductor wafer. It can provide non-destructive quantitative measurements of surface roughness and geometric pattern characterization with nanometric axial resolution in real-time. However, the measurement results may be affected by an additional phase curvature introduced by the microscope objective (MO) used in DHM. It needs to be removed either by numerical compensation or by physical compensation.We present a method of physical spherical phase compensation for reflection DHM in the Michelson configuration. In the object arm, collimated light is used for illumination. Due to the use of the MO, the object wavefront may have a spherical phase curvature. In the reference arm, a lens and mirror combination is used to generate a spherical recording reference wave in order to physically compensate the spherical phase curvature of the object wavefront. By controlling the position of the mirror and the sample stage, the compensation process has been demonstrated. The relative positions of the test specimen and the reference mirror must be fixed for the physical spherical phase to be totally compensated. A numerical plane reference wave is preferred for the numerical reconstruction of the phase introduced by the test specimen. Experimental results on wafer pattern recognition are also given.     

共轭读出法消除光致变色材料全息记录中的相位畸变

采用有机光致变色材料-吲哚俘精酰胺/PMMA薄膜作为可擦写全息记录介质,建立了一种能消除全息光存储中相位畸变的光路.此光路采用参考光的相位共轭光再现全息图,能有效消除物光路中由于光学元件失调和记录材料缺陷引起的相位畸变,是全息光存储中提高再现像像质的一种有用光路.     

Determining the complex point-spread function of a scanning differential heterodyne microscope

For a scanning differential heterodyne microscope, the two-dimensional point spread function and the pupil function are determined from experimental measurements of the intensity distribution of a probing beam. The phase components of these functions were restored from the measured distributions using the Gerchberg-Saxton algorithm. Based on the two-dimensional pupil function, the three-dimensional point-spread function is determined in the scalar approximation using the Debye integral. The aberration function of the microscope is analyzed from the viewpoint of the composition of aberrations and method of their balancing. The focusing adjustment criterion of the microscope for an object under study by the phase response from the test structure is considered.     

Wavefront sensing with random amplitude mask and phase retrieval

A light beam with an ideal wavefront that is transmitted or reflected from an object is modified by different characteristics of the object such as shape, refractive index, density, or temperature. Wavefront sensing therefore yields valuable information about the system or the changes happening to the system. A new method for wavefront sensing using a random amplitude mask and a phase retrieval method based on the Rayleigh-Sommerfeld wave propagation equation is described. The proposed method has many potential applications ranging from phase contrast imaging and measurement of lens aberration to shape measurement of three-dimensional objects.     

Non-linear aberration of a self-focussing electromagnetic wave beam

In this paper the aberrations of a centered axially symmetric electromagnetic wave beam in a nonlinear medium are examined. A formula is obtained, which describes the intensity distribution of the beam near the aberration point. The procedure for investigating high-order aberrations is demonstrated. The influence of the initial distribution of the intensity and the phase on the picture of the aberration is examined.     

Phase aberration compensation of digital holographic microscopy based on least squares surface fitting

Digital holographic microscopy allows the numerical reconstruction of the complex wavefront of samples, especially biological samples such as living cells. In digital holographic microscopy, a microscope objective is introduced to improve the transverse resolution of the sample; however a phase aberration in the object wavefront is also brought along, which will affect the phase distribution of the reconstructed image. We propose here a numerical method to compensate for the phase aberration of thin transparent objects with a single hologram. The least squares surface fitting with points number less than the matrix of the original hologram is performed on the unwrapped phase distribution to remove the unwanted wavefront curvature. The proposed method is demonstrated with the samples of the cicada wings and epidermal cells of garlic, and the experimental results are consistent with that of the double exposure method.     

Study on aberration suppressing methods in digital micro-holography

This paper discusses methods to eliminate and/or suppress aberrations in digital micro-holography (DMH) with specific examples for post-magnification DMH and lens-less DMH along with a phase subtraction approach. In post-magnification DMH, the reference wave interferes with the object wave to form a hologram, which is then magnified by a microscope objective (MO) and recorded by CCD. This compensates for the quadratic phase error usually present in the pre-magnification DMH and also it is easier to isolate the useful information in pre-processing. In this paper differences between post-magnification and pre-magnification DMH are compared and analyzed with the specimen of a phase grating with period 30 lines/mm and height 0.3 μm. A lens-less DMH is also described that can eliminate the quadratic error and most of wave-front errors of the projecting light. Finally, a phase subtraction method is demonstrated which can eliminate most of the system aberrations.     

A two-stage method to correct aberrations induced by slide slant in bright-field microscopy

To achieve optimal image quality in bright field microscopy, the slide surface should be perpendicular to the optical axis of the microscope. However, in the recently proposed “slanted scan” slide acquisition technique, scan speed is increased by purposely slanting the slide by a small angle (of 3–5°) so that multiple focal depths can be imaged simultaneously. In this case, the slanted slide introduces a bend in the point spread function (PSF), resulting in a coma and other aberrations that degrade image quality. In this paper, we propose a two-stage deconvolution method specifically designed to correct the aberrations induced by a slanted scan, but with general applicability to high-resolution bright-field microscopy. Specifically, we initially apply phase deconvolution to correct the dominating coma aberration, before applying a conventional semi-blind deconvolution method to further improve image resolution and contrast. We also propose a novel method to estimate the degree of coma aberration and the PSF of the optics utilising actual cytology specimens. The efficacy of the proposed algorithm is demonstrated quantitatively on simulated data, against a ground-truth (object) image, and qualitatively on cervical cytology specimens. Results demonstrate both improved convergence speed of the two-stage approach, especially when correcting the bend in the PSF, and a resultant image quality that is comparable to a conventionally (flat) scanned specimen.     

Quantitative phase-contrast microscopy by a lateral shear approach to digital holographic image reconstruction

Combining the concept of lateral shear interferometry (LSI) within a digital holography microscope, we demonstrate that it is possible to obtain quantitative optical phase measurement in microscopy by a new single-image-processing procedure. Numerical lateral shear of the reconstructed wavefront in the image plane makes it possible to retrieve the derivative of the wavefront and remove the defocus aberration term introduced by the microscope objective. The method is tested to investigate a silicon structure and a mouse cell line.     

Phase plates for wave-aberration compensation in the human eye

We present a method for manufacturing phase plates to compensate for the wave aberration in the human eye. The wave aberration of the eye is measured in vivo by a new laser ray-tracing method and then compensated for by a phase plate placed in front of the eye. This plate is made from a gray-level single-mask photosculpture in photoresist. Two experiments were carried out, first with an artificial eye and then with a human eye: 80% compensation for the wave aberration was achieved in both cases.     

应用波像差理论计算Kirkpatrick-Baez系统的调制传递函数

调制传递函数（modulation transfer function,MTF）曲线可以客观地评价光学系统的成像性能。基于平面对称光学系统的波像差理论,给出了Kirkpatrick-Baez（KB）显微镜光学系统主要像差的波像差计算表达式,采用自相关法,通过Gauss-Legendre数值积分求解并绘制了KB系统的MTF曲线,且与光学分析软件的MTF曲线进行对比。结果表明:应用波像差理论计算调制传递函数不仅可以定量直观地分析KB系统的成像性能,而且能进一步解析光学系统的单项像差分布情况,提供更具针对性的系统优化方案。     

Visualization of phase conjugate ultrasound waves passed through inhomogeneous layer

Compensation of phase distortions of ultrasound beams by means of parametric phase conjugation is visualized. Quasi-plane and focused primary beams were distorted by a polymer aberration layer introduced between the primary wave source and the wave phase conjugator. It is demonstrated acousto-optically that, while the acoustic field is strongly irregular in the area between aberration layer and conjugator, the phase conjugate wave visibly reproduces the primary beams in the area between the layer and the primary wave source. The phenomenon is observed in supercritical mode of parametric amplification when intensity of phase conjugate wave is high enough for manifestations of acoustic nonlinearities in water.     

无透镜傅里叶变换数字全息术中非共面误差的自动补偿算法

无透镜傅里叶变换数字全息术对活细胞进行显微相衬成像测量时,由于细胞处于运动状态导致记录物体与参考点源难以共面,从而引入成像误差.本文提出了一种有效可靠的基于位相分布的自动对焦再现算法,只需通过一幅全息图,即可确定出记录距离的优化数值解,并结合一次位相线性拟合,成功补偿掉非共面误差.将这种算法应用到活体宫颈癌细胞的相衬成像实验中获得了较为理想的实验结果,表明了这种自动补偿非共面误差算法的可行性. 关键词： 数字全息 无透镜傅里叶变换全息 自动对焦 位相畸变校正     

HREM of the [111] surfaces of iron oxide nanoparticles

Mixed phase Fe3O4-gamma-Fe2O3 (magnetite-maghemite) iron oxide nanoparticles have been fabricated by colloidal routes. HRTEM/HRSTEM images of the nanoparticles show the presence of [111] facets that terminate with enhanced contrast, which is shown to be caused by the presence of additional cations at the edges of the nanoparticles. HRTEM images were taken on a FEI CM200 FEGTEM, a JEOL 3100 with a LaB6 source, and a double aberration corrected JEOL-JEM 2200FS FEGTEM. The enhanced contrast effect was observed on the [111] surface atomic layers resolved using each machine. HRSTEM images, taken on an aberration corrected STEM, resolved enhanced contrast at specific surface sites. Exit wave reconstruction was also carried out on focal series taken on a double aberration corrected JEOL-JEM 2200FS and showed similar highly resolved enhanced contrast at specific surface cation sites. It is apparent that additional cations are occupying the [111] terminating layers of these nanoparticle surfaces. The use of different microscopes and techniques in this paper provides strong evidence that the enhanced contrast is a real effect and not an effect caused by microscope aberrations.     

Wave aberration function and its definition

A definition of a wave aberration as a phase shift upon composition of light waves in the image of a point is given using the concept of point eikonal. An expression that determines the total differential of a wave aberration function is obtained and the condition of its integrability is determined. The sequence of the wave aberration function definition at the known functions of the meridional and sagittal components of lateral aberration is presented.     

Wave field restoration using focal-depth extension techniques under dynamic hollow-cone illumination

Wave field restoration has been achieved based on focal-depth extension under dynamic hollow-cone illumination using an optical microscope. Two kinds of focal-depth extension techniques, such as moving focus averaging and an objective annular aperture, were compared and their effectiveness for transmission electron microscopy applications is discussed. Both techniques enabled resolution enhancement at the same level and effectively obtained aberration-free phase and amplitude images separately by canceling out the rotationally invariant component of system wave aberrations, such as spherical aberration. The influence of nonlinear interference on image contrast was confirmed as significant by using a figure 8 shaped spatial frequency filter.     

Measuring lateral efficiency of optical traps: The effect of tube length

An optical tweezer setup is made based on a custom-designed inverted microscope, which can work both in finite and infinite tube length microscopy modes. It is shown that the spherical aberration due to the mismatch in the refractive indices of the specimen (water) and the immersion oil as well as the wavelength can be partially compensated by introducing another source for the spherical aberration provided it has the opposite sign. Changing the tube length is shown to be a good candidate for this effect: an improvement of up to a factor of four has been observed in the lateral efficiency of the trap.     

光束斜入射的折反射组合光学系统的像差计算

基于平面对称光栅反射镜系统的像差理论,根据光学元件材料折射率对成像光束波像差进行修正,将像差理论扩展到平面对称折射光学系统,使得该理论可应用于光束斜入射的折反射组合光学系统的像差计算;指出在折反射组合光学系统中考虑物像方空间折射率的关系,以及入射角方向的定义,使得波像差的计算表达式保持统一;应用扩展后的像差理论对折反射...     

Non-contact acoustic method for the simultaneous measurement of thickness and acoustic properties of biological tissues

The frequency dependence of the magnitude and phase shift of reflected waves from a thin tissue specimen has been found to yield characteristics patterns as a function of the specimen thickness. By analysis of those characteristic patterns, the specimen thickness can be determined so that the attenuation constant and the sound speed can be obtained by a non-contact procedure. The method is demonstrated in the frequency range 100 MHz to 200 MHz, using a scanning acoustic microscope and approximately 10 μm thick myocardial tissue samples of human origin, one paraffin embedded.