We demonstrate two methods based on Fourier plane filtering using (a) a fractional spiral phase plate (SPP) and (b) an off-axial SPP for phase contrast enhancement in optical microscopy. In comparison to previous works, a spatially incoherent LED is used in the Köhler illumination as the light source to illuminate the biological specimen. We demonstrate that both these methods can transform the phase specimen into a relief-like view even under such illumination. The degree and orientation of enhancement can be controlled by changing the phase structure of the filter. The SPP is displayed on a phase-only spatial light modulator, and can be integrated into the optical path of standard microscopes. 相似文献
Low‐coherence optical microscopy or optical coherence microscopy uses light with short coherence length. The well‐known case is: “white‐light interferometry”, which became recently more known as: “optical coherence tomography”. However, when lenses and microscope objectives are used to create interferometric images, in what is known classically as “interference microscopy” or today as “full‐field optical coherence tomography” the spatial coherence starts to play a critical role. In this article the coherence effects in low‐coherence optical microscopy are reviewed. As this technology is becoming increasingly publicized due to its importance in three‐dimensional imaging, particularly of scattering biological media and optical metrology, the understanding of the fundamental physics behind it is essential. The interplay between longitudinal spatial coherence and temporal coherence and the effects associated with them are discussed in detail particularly when high numerical apertures are used. An important conclusion of this study is that a high‐contrast, high‐resolution system for imaging of multilayered samples is the one that uses narrowband illumination and high‐NA objectives with an index‐matching fluid. Such a system, when combined with frequency‐domain operation, can reveal nearly real‐time three‐dimensional images, and is thus competitive with confocal microscopy. 相似文献
The advent of computer-generated or synthetic holography has created a wealth of possibilities for wavefront shaping in optics. We discuss the impact this has had on optical microscopy. Synthetic Holographic Microscopy utilises wavefront shaping by a computer-generated ‘hologram’ (CGH) to modify light on the illumination or the detection side, or both. This enables modifications of the general sample appearance concerning contrast, resolution and other aspects. Multiplexing CGHs can perform several tasks at once, for instance splitting the image into sub-images corresponding to different depths in the sample, or displaying differently contrasted images of the sample, e.g. bright field, darkfield or (spiral) phase contrast, in different sub-images. We give an overview of the options and discuss the advantages and disadvantages of using programmable holographic elements inside an optical microscope. 相似文献
Recently it has been demonstrated that spatial filtering of images in microscopy with a spiral phase element in a Fourier plane of the optical path results in a strong edge enhancement of object structures. In principle the operation is isotropic, i.e., all phase edges of a sample object are highlighted simultaneously, independent of their local direction. However, here we demonstrate that the symmetry can be broken intentionally by controlling the phase of the central area of a spiral phase hologram, which is displayed at a computer controlled spatial light modulator. This produces an apparent shadow effect which can be rotated at video rate. The resulting relieflike impression of the sample topography with a longitudinal resolution in the subwavelength regime is demonstrated by imaging a standard low contrast test sample consisting of a human cheek cell. 相似文献
Novel X‐ray imaging of structural domains in a ferroelectric epitaxial thin film using diffraction contrast is presented. The full‐field hard X‐ray microscope uses the surface scattering signal, in a reflectivity or diffraction experiment, to spatially resolve the local structure with 70 nm lateral spatial resolution and sub‐nanometer height sensitivity. Sub‐second X‐ray exposures can be used to acquire a 14 µm × 14 µm image with an effective pixel size of 20 nm on the sample. The optical configuration and various engineering considerations that are necessary to achieve optimal imaging resolution and contrast in this type of microscopy are discussed. 相似文献
X‐ray microscopy is a commonly used method especially in material science application, where the large penetration depth of X‐rays is necessary for three‐dimensional structural studies of thick specimens with high‐Z elements. In this paper it is shown that full‐field X‐ray microscopy at 6.2 keV can be utilized for imaging of biological specimens with high resolution. A full‐field Zernike phase‐contrast microscope based on diffractive optics is used to study lipid droplet formation in hepatoma cells. It is shown that the contrast of the images is comparable with that of electron microscopy, and even better contrast at tender X‐ray energies between 2.5 keV and 4 keV is expected. 相似文献
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. 相似文献
Photoacoustic microscopy (PAM) is a hybrid in vivo imaging technique that acoustically detects optical contrast via the photoacoustic effect. Unlike pure optical microscopic techniques, PAM takes advantage of the weak acoustic scattering in tissue and thus breaks through the optical diffusion limit (∼1 mm in soft tissue). With its excellent scalability, PAM can provide high‐resolution images at desired maximum imaging depths up to a few millimeters. Compared with backscattering‐based confocal microscopy and optical coherence tomography, PAM provides absorption contrast instead of scattering contrast. Furthermore, PAM can image more molecules, endogenous or exogenous, at their absorbing wavelengths than fluorescence‐based methods, such as wide‐field, confocal, and multi‐photon microscopy. Most importantly, PAM can simultaneously image anatomical, functional, molecular, flow dynamic and metabolic contrasts in vivo. Focusing on state‐of‐the‐art developments in PAM, this Review discusses the key features of PAM implementations and their applications in biomedical studies. 相似文献
We present a spiral phase filtering system with a large tolerance for edge enhancement of both phase and amplitude objects in optical microscopy.The method is based on a Fourier 4-f spatial filtering system.A phase mismatched spiral phase plate (SPP) fabricated by electron beam lithography is employed as the radial Hilbert transform for image edge enhancement.Compared with holography,SPP is simple,economical,reliable,and easy to integrate. 相似文献
This paper briefly reviews the basics of optical Fourier techniques (OFT) and applications for medical image processing as well as phase contrast imaging of live biological specimens. Enhancement of microcalcifications in a mammogram for early diagnosis of breast cancer is the main focus. Various spatial filtering techniques such as conventional 4f filtering using a spatial mask, photoinduced polarization rotation in photosensitive materials, Fourier holography, and nonlinear transmission characteristics of optical materials are discussed for processing mammograms. We also reviewed how the intensity dependent refractive index can be exploited as a phase filter for phase contrast imaging with a coherent source. This novel approach represents a significant advance in phase contrast microscopy. 相似文献
Dark-field illumination is known to enhance scattering contrast in optical microscopy. We combined this concept with Fourier domain optical coherence microscopy (OCM). The detection and illumination paths are decoupled, and only the scattered light originating from the sample generates the tomogram signal, whereas any specular reflection is highly suppressed. We analyze and discuss this dark-field OCM concept and present its superior imaging quality on live cell samples. 相似文献
As a revolutionary observation tool in life science, biomedical, and material science, optical microscopy allows imaging of samples with high spatial resolution and a wide field of view. However, conventional microscopy methods are limited to single imaging and cannot accomplish real-time image processing. The edge detection, image enhancement and phase visualization schemes have attracted great interest with the rapid development of optical analog computing. The two main physical mechanisms that enable optical analog computing originate from two geometric phases: the spin-redirection Rytov-Vlasimirskii-Berry (RVB) phase and the Pancharatnam-Berry (PB) phase. Here, we review the basic principles and recent research progress of the RVB phase and PB phase based optical differentiators. Then we focus on the innovative and emerging applications of optical analog computing in microscopic imaging. Optical analog computing is accelerating the transformation of information processing from classical imaging to quantum techniques. Its intersection with optical microscopy opens opportunities for the development of versatile and compact optical microscopy systems. 相似文献
The spatial light modulators (SLMs) at the Fourier planes of a conventional joint-transform correlation (JTC) are eliminated using simple grating filters such as rectangular and triangular ones. To achieve this goal, two non-conventional real-time JTC architectures are proposed: (a) the grating filter is used along with a heterodyning and one-dimensional optical scanning technique to capture the cross-correlation functions of the input images without major processing and (b) the one-dimensional optical scanning is eliminated to achieve a faster (but a little bit more complicated) processing. The proposed techniques significantly reduce the time processing needed for real-time applications by eliminating the drawbacks of the non-ideal characteristics of the SLMs. 相似文献
This article reviews the state of the art of ultrafast transient absorption microscopy, discusses current experimental concepts and highlights future challenges. The advantages of transient absorption microscopy over other micro‐spectroscopic techniques are its high optical resolution combined with high temporal resolution as well as its ability to study non‐fluorescent and weakly fluorescent molecular species and to probe excited‐state processes. In conventional transient absorption spectroscopy the spectroscopic information usually presents a spatial average over the focal spot of the typically weakly focused probe beam. Transient absorption microscopy, however, enables investigations of the excited state dynamics in individual microscopic areas of a sample. Hence, the technique does not only yield detailed morphological information based on a label‐free molecular contrast, but also gives insight into the ultrafast morphology‐dependent photoinduced processes in heterogeneous samples. Different variations of transient absorption microscopy have found a number of applications ranging from material sciences to biology, which are discussed in this review together with different setup modifications and approaches towards transient absorption spectroscopy with spatial resolution below the diffraction limit.