Emulated super-resolution using quantitative phase microscopy

In the case of coherent illumination, knowledge of the phase and the amplitude of a light wave constitutes complete information. Phase and amplitude information can now be simply acquired using the technique of quantitative phase microscopy. It has been shown that this information allows other imaging modalities to be emulated. In this paper we consider how this information may be used to perform a form of super-resolution by emulating the effect of an annular pupil.     

Quantitative phase retrieval in X‐ray Zernike phase contrast microscopy

In recent years, increasing attention has been devoted to X‐ray phase contrast imaging, since it can provide high‐contrast images by using phase variations. Among the different existing techniques, Zernike phase contrast microscopy is one of the most popular phase‐sensitive techniques for investigating the fine structure of the sample at high spatial resolution. In X‐ray Zernike phase contrast microscopy, the image contrast is indeed a mixture of absorption and phase contrast. Therefore, this technique just provides qualitative information on the object, which makes the interpretation of the image difficult. In this contribution, an approach is proposed for quantitative phase retrieval in X‐ray Zernike phase contrast microscopy. By shifting the phase of the direct light by π/2 and 3π/2, two images of the same object are measured successively. The phase information of the object can then be quantitatively retrieved by a proper combination of the measured images. Numerical experiments were carried out and the results confirmed the feasibility of the proposed method. It is expected that the proposed method will find widespread applications in biology, materials science and so on.     

A new method for information retrieval in two-dimensional grating-based X-ray phase contrast imaging

Grating-based X-ray phase contrast imaging has been demonstrated to be an extremely powerful phase-sensitive imaging technique.By using two-dimensional(2D) gratings,the observable contrast is extended to two refraction directions.Recently,we have developed a novel reverse-projection(RP) method,which is capable of retrieving the object information efficiently with one-dimensional(1D) grating-based phase contrast imaging.In this contribution,we present its extension to the 2D grating-based X-ray phase contrast imaging,named the two-dimensional reverseprojection(2D-RP) method,for information retrieval.The method takes into account the nonlinear contributions of two refraction directions and allows the retrieval of the absorption,the horizontal and the vertical refraction images.The obtained information can be used for the reconstruction of the three-dimensional phase gradient field,and for an improved phase map retrieval and reconstruction.Numerical experiments are carried out,and the results confirm the validity of the 2D-RP method.     

Zernike phase contrast in scanning microscopy with X-rays

Scanning X-ray microscopy focuses radiation to a small spot and probes the sample by raster scanning. It allows information to be obtained from secondary signals such as X-ray fluorescence, which yields an elemental mapping of the sample not available in full-field imaging. The analysis and interpretation from these secondary signals can be considerably enhanced if these data are coupled with structural information from transmission imaging. However, absorption often is negligible and phase contrast has not been easily available. Originally introduced with visible light, Zernike phase contrast(1) is a well-established technique in full-field X-ray microscopes for visualization of weakly absorbing samples(2-7). On the basis of reciprocity, we demonstrate the implementation of Zernike phase contrast in scanning X-ray microscopy, revealing structural detail simultaneously with hard-X-ray trace-element measurements. The method is straightforward to implement without significant influence on the resolution of the fluorescence images and delivers complementary information. We show images of biological specimens that clearly demonstrate the advantage of correlating morphology with elemental information.     

Confocal diffraction phase microscopy of live cells

We present a new quantitative phase microscopy technique, confocal diffraction phase microscopy, which provides quantitative phase measurements from localized sites on a sample with high sensitivity. The technique combines common-path interferometry with confocal microscopy in a transmission geometry. The capability of the technique for static imaging is demonstrated by imaging polystyrene microspheres and live HT29 cells, while dynamic imaging is demonstrated by quantifying the nanometer scale fluctuations of red blood cell membranes.     

Dynamic range improvement in NMR imaging using phase scrambling

Data collection efficiency in NMR imaging is impaired if the dynamic range of the receiver system is limited in comparison with that of the observed signal. This situation may occur in high-resolution proton imaging of large objects at high magnetic field strengths. The efficiency with which information is received can be increased by reducing the peak amplitude of the spin response by varying the phase distribution of the excited spins. This phase scrambling technique may be implemented using tailored RF excitation or by dephasing using nonlinear magnetic field gradients and can be applied in all dimensions of an acquired data set, providing a significant reduction in the dynamic range requirements of the detection electronics. Experimental results using 2D Fourier imaging have obtained up to 25 dB reduction in peak signal intensities. Image signal-to-noise ratios improved up to a factor of 6, with actual values dependent on experimental conditions. Simulation studies show that computational noise introduced during Fourier transformation is significantly reduced when phase scrambling is employed.     

Spectral-domain optical coherence phase microscopy for quantitative phase-contrast imaging

We describe a novel microscopy technique for quantitative phase-contrast imaging of a transparent specimen. The technique is based on depth-resolved phase information provided by common path spectral-domain optical coherence tomography and can measure minute phase variations caused by changes in refractive index and thickness inside the specimen. We demonstrate subnanometer level path-length sensitivity and present images obtained on reflection from a known phase object and human epithelial cheek cells.     

基于单一分光棱镜干涉仪的双通路定量相位显微术

仅仅使用一个单独的分光棱镜(BS),实现了一种用于生物细胞三维成像的双通路定量相位显微术.不同于传统的使用方法,将BS倾斜放置,使中央半反射层与入射光光轴之间存在一个非常小的角度.这样基于BS的分光特性,经过BS后的透射光束和反射光束将会叠加在一起并形成干涉.调节样品位置,利用相机拍摄同时获得了存在π相移的双通路干涉图.这种离轴干涉模式,只需要记录单幅干涉图就可以获得真实的相位信息,方法结构简单,易于操作,适用于微小透明样品的三维形貌测量.     

联合谱域与深度域光谱相位显微方法

提出了一种联合谱域与深度域光谱相位显微方法, 该方法利用谱域相位信息克服2π歧义, 并结合深度域相位信息, 以实现高动态范围、高灵敏度的相位检测. 首先通过理论推导和信号模拟, 进行了深度域相位和谱域相位的灵敏度比较, 证明了深度域相位在灵敏度上要高于谱域相位. 进而详细介绍了联合谱域与深度域光谱相位显微方法. 最后通过盖玻片和光学分辨率板实验验证了所提出的联合谱域 与深度域光谱相位显微方法能够在实现高动态测量范围的同时保持高相位灵敏度. 关键词： 光谱相位显微方法 动态范围 灵敏度     

Extended depth of focus in tomographic phase microscopy using a propagation algorithm

Tomographic phase microscopy is a laser interferometry technique in which a 3D refractive index map of a biological sample is constructed from quantitative phase images collected at a set of illumination angles. Although the resulting tomographic images provide valuable information, their resolution declines at axial distances beyond about 1 microm from the focal plane. We describe an improved 3D reconstruction algorithm in which the field at the focal plane is numerically propagated to depths throughout the sample. Diffraction is thus incorporated, extending the depth of focus to more than 10 mum. Tomograms with improved focal depth are demonstrated for single HT29 cells.     

Phase-sensitive fat suppression steady-state free procession sequence with phase correction

Robust and fast fat suppression is a challenge in balanced steady-state free precession （SSFP） magnetic resonance imaging. Although single-acquisition phase-sensitive SSFP can provide fat-suppressed images in short scan time, phase errors, especially spatially-dependent phase shift, caused by a variety of factors may result in misplacement of fat and water voxels. In this paper, a novel phase correction algorithm was used to calibrate those phase errors during image reconstruction. This algorithm corrects phase by region growing, employing both the magnitude and the phase information of image pixels. Phantom and in vivo imagings were performed to validate the technique. As a result, excellent fat-suppressed images were acquired by using single-acquisition phase-sensitive SSFP with phase correction.     

Phase-sensitive fat suppression steady-state free procession sequence with phase correction

Robust and fast fat suppression is a challenge in balanced steady-state free precession (SSFP) magnetic resonance imaging. Although single-acquisition phase-sensitive SSFP can provide fat-suppressed images in short scan time, phase errors, especially spatially-dependent phase shift, caused by a variety of factors may result in misplacement of fat and water voxels. In this paper, a novel phase correction algorithm was used to calibrate those phase errors during image reconstruction. This algorithm corrects phase by region growing, employing both the magnitude and the phase information of image pixels. Phantom and \textit{in vivo} imagings were performed to validate the technique. As a result, excellent fat-suppressed images were acquired by using single-acquisition phase-sensitive SSFP with phase correction.     

Dual-wavelength linear regression phase unwrapping in three-dimensional microscopic images of cancer cells

We present a study of the three-dimensional structure of cancer cells using dual-wavelength phase-imaging digital holographic microscopy. Phase imaging of objects with optical height variation greater than the wavelength of light is ambiguous and causes phase wrapping. By comparing two phase images recorded at different wavelengths, the images can be accurately unwrapped. The unwrapping method is computationally fast and straightforward, and it can process complex topologies. Additionally, the limitations on the total optical height are significantly relaxed. This new methodology is widely applicable to other phase-imaging techniques as well as in applications beyond optical microscopy.     

Jones phase microscopy of transparent and anisotropic samples

We developed an interferometric microscopy technique, referred to as Jones phase microscopy, capable of extracting the spatially resolved Jones polarization matrix associated with transparent and anisotropic samples. This is a generalization of quantitative phase imaging, which is recovered from one diagonal element of the measured matrix. The principle of the technique is demonstrated with measurements of a liquid crystal spatial light modulator and the potential for live cell imaging with experiments on live neurons in culture.     

Compact,common path quantitative phase microscopic techniques for imaging cell dynamics

Microscopy using visible electromagnetic radiation can be used to investigate living cells in various environments. But bright field microscopy only provides two-dimensional (2D) intensity distribution at a single object plane. One of the ways to retrieve object height/thickness information is to employ quantitative phase microscopic (QPM) techniques. Interferometric QPM techniques are widely used for this. Digital holographic microscopy (DHM) is one of the state-of-the-art methods for quantitative three-dimensional (3D) imaging. Usually it is implemented in two-beam geometry, which is prone to mechanical vibrations. But to study dynamics of objects like red blood cells, one needs temporal stability much better than the fluctuations of the object, which the two-beam geometry fails to deliver. One way to overcome this hurdle is to use self-referencing techniques, in which a portion of the object beam will act as the reference beam. Here the development of self-referencing QPM techniques is described along with the results.     

Three-dimensional color photon counting microscopy using Bayesian estimation with adaptive priori information

In this Letter, we propose a novel three-dimensional(3D) color microscopy for microorganisms under photonstarved conditions using photon counting integral imaging and Bayesian estimation with adaptive priori information. In photon counting integral imaging, 3D images can be visualized using maximum likelihood estimation(MLE). However, since MLE does not consider a priori information of objects, the visual quality of 3D images may not be accurate. In addition, the only grayscale image can be reconstructed. Therefore, to enhance the visual quality of 3D images, we propose photon counting microscopy using maximum a posteriori with adaptive priori information. In addition, we consider a wavelength of each basic color channel to reconstruct 3D color images. To verify our proposed method, we carry out optical experiments.     

X射线相位衬度成像

文章使用形象、生动、通俗、易懂的语言,介绍X射线相位衬度成像的基本概念,物理思想和方法,其中包括X射线的基本性质、光的波粒二象性、同步辐射X射线光源和常规非相干X射线光源的相位相干性,以及X射线相位衬度成像方法、三维成像的基本原理和相位衬度成像的最新进展,将抽象的相位、相位一阶导数和相位二阶导数概念与形象的光波阵面平移、倾斜和弯曲等形变联系在一起,着重介绍相位衬度成像发展中的创新思想,力图使读者能分享人类文明在这个学科发展中积累的精神财富.     

X射线相位衬度成像

文章使用形象、生动、通俗、易懂的语言,介绍X射线相位衬度成像的基本概念,物理思想和方法,其中包括X射线的基本性质、光的波粒二象性、同步辐射X射线光源和常规非相干X射线光源的相位相干性,以及X射线相位衬度成像方法、三维成像的基本原理和相位衬度成像的最新进展,将抽象的相位、相位一阶导数和相位二阶导数概念与形象的光波阵面平移、倾斜和弯曲等形变联系在一起,着重介绍相位衬度成像发展中的创新思想,力图使读者能分享人类文明在这个学科发展中积累的精神财富.     

Diffraction phase microscopy for quantifying cell structure and dynamics

We have developed diffraction phase microscopy as a new technique for quantitative phase imaging of biological structures. The method combines the principles of common path interferometry and single-shot phase imaging and is characterized by subnanometer path-length stability and millisecond-scale acquisition time. The potential of the technique for quantifying nanoscale motions in live cells is demonstrated by experiments on red blood cells.     

SNR phase order k-space encoding (SPOKE)

A method of determining the phase-encode order for MR Fourier-encoded imaging is described, which provides an additional option for optimizing images from samples with signals that change during data acquisition. Examples are in hyperpolarized helium gas imaging of the lungs where polarization is lost with each RF pulse or the signal changes observed in rapid dynamic studies with T₁ or T₂* contrast agents when mixing is taking place. The method uses a single frequency-encoded projection in the proposed phase-encoding direction. The projection is subsequently sorted into signal-to-noise ratio (SNR) order. The indices of the sorted array are then used to create the phase-encode table to be used for the scan. This phase table is sorted in descending SNR order for signals that decrease during data acquisition and in ascending order for signals that increase during data acquisition. Simulations suggest that this technique can produce higher resolution than centric-ordered phase encoding at the expense of increased modulation (ghosting) artifact for dynamically changing signals. Initial practical implementation of the technique has been carried out on a dedicated 0.2-T Niche MR system, and the test object results agree well with simulations. Hyperpolarized 3-He lung images have also been acquired and postprocessed using the SNR phase order k-space encoding (SPOKE) methodology and show potential for improved imaging with high flip angles where polarization is rapidly lost. Applications may also be found for 3D volumetric acquisitions where two dimensions can be SPOKE encoded.