RESUMEN
The characterization of the structure of highly hierarchical biosamples such as collagen-based tissues at the scale of tens of nanometers is essential to correlate the tissue structure with its growth processes. Coherent x-ray Bragg ptychography is an innovative imaging technique that gives high resolution images of the ordered parts of such samples. Herein, we report how we used this method to image the collagen fibrillar ultrastructure of intact rat tail tendons. The images show ordered fibrils extending over 10-20 µm in length, with a quantifiable D-banding spacing variation of 0.2%. Occasional defects in the fibrils distribution have also been observed, likely indicating fibrillar fusion events.
Asunto(s)
Colágenos Fibrilares/metabolismo , Imagen Molecular/métodos , Tendones/metabolismo , Algoritmos , Animales , Procesamiento de Imagen Asistido por Computador , Ratas , Difracción de Rayos X , Rayos XRESUMEN
Accurate knowledge of translation positions is essential in ptychography to achieve a good image quality and the diffraction limited resolution. We propose a method to retrieve and correct position errors during the image reconstruction iterations. Sub-pixel position accuracy after refinement is shown to be achievable within several tens of iterations. Simulation and experimental results for both optical and X-ray wavelengths are given. The method improves both the quality of the retrieved object image and relaxes the position accuracy requirement while acquiring the diffraction patterns.
RESUMEN
In this paper the coherence requirement for different holographic setups (Fresnel hologram, Fourier hologram, and image-plane hologram) is compared. This analysis is based on the investigation of the recorded interference pattern from the superposition of reference wave and object wave in in-line and off-axis mode. The outcome of this investigation can support the choice of light source needed for certain digital holographic setups, as well as the selection of the best applicable setup to take advantage of new short coherence light sources. Moreover, as a byproduct of this investigation, the minimum required recording distance (focal length) to enable Nyquist sampling of the recorded hologram is obtained.
RESUMEN
This paper shows that visible-light ptychography can be used to distinguish quantitatively between healthy and tumorous unstained cells. Advantages of ptychography in comparison to conventional phase-sensitive imaging techniques are highlighted. A novel procedure to automatically refocus ptychographic reconstructions is also presented, which improves quantitative analysis.
Asunto(s)
Neoplasias Encefálicas/patología , Glioma/patología , Interpretación de Imagen Asistida por Computador/métodos , Microscopía de Contraste de Fase/métodos , Nefelometría y Turbidimetría/métodos , Refractometría/métodos , Animales , Diagnóstico Diferencial , RatasRESUMEN
Coherent diffractive imaging of objects is made considerably more practicable by using ptychography, where a set of diffraction patterns replaces a single measurement and introduces a high degree of redundancy into the recorded data. Here we demonstrate that this redundancy allows diffraction patterns to be extrapolated beyond the aperture of the recording device, leading to superresolved images, improving the limit on the finest feature separation by more than a factor of 3.
RESUMEN
Most implementations of ptychography on the electron microscope operate in scanning transmission (STEM) mode, where a small focussed probe beam is rapidly scanned across the sample. In this paper we introduce a different approach based on near-field ptychography, where the focussed beam is replaced by a wide-field, structured illumination, realised through a purpose-designed etched Silicon Nitride window. We show that fields of view as large as 100 µm2 can be imaged using the new approach, and that quantitative electron phase images can be reconstructed from as few as nine near-field diffraction pattern measurements.
RESUMEN
Quantitative phase microscopy offers a range of benefits over conventional phase-contrast techniques. For example, changes in refractive index and specimen thickness can be extrapolated and images can be refocused subsequent to their recording. In this Letter, we detail a lensless, quantitative phase microscope with a wide field of view and a useful resolution. The microscope uses the recently reported coherent diffractive imaging technique of ptychography to generate images from recorded diffraction patterns.
Asunto(s)
Microscopía de Interferencia/métodos , Microscopía de Contraste de Fase/métodos , Óptica y Fotónica , Algoritmos , Línea Celular Tumoral , Diseño de Equipo , Holografía , Humanos , Procesamiento de Imagen Asistido por Computador , Lentes , Refractometría , Reproducibilidad de los Resultados , SemiconductoresRESUMEN
Ptychographic iterative engine (PIE), a form of coherent diffractive imaging in which the phase of both low-angle and high-angle diffraction data is used to form an image without a lens, can in principle deliver wavelength-limited resolution. Working at low accelerating voltages (30 keV) reduces specimen knock-on damage, increases high-resolution contrast and allows a scanning electron microscope to be used as a transmission electron microscopy. However, electrons are very strongly scattered at low energy. To that end, the effect of dynamical scattering on PIE at low accelerating voltages is calculated to illustrate the possibility of generating high-resolution images with low energy electrons.
RESUMEN
We investigate a strategy for separating the influence of three-dimensional scattering effects in tilt-series reconstruction, a method for computationally increasing the resolution of a transmission microscope with an objective lens of small numerical aperture, as occurs in the transmission electron microscope (TEM). Recent work with visible light refers to the method as Fourier ptychography. To date, reconstruction methods presume that the object is thin enough so that the beam tilt induces only a shift of the diffraction pattern in the back focal plane. In fact, it is well known that the diffraction pattern changes as a function of beam tilt when the object is thick. In this paper, we use a simple visible light model to demonstrate a proof-of-principle of a new reconstruction algorithm that can cope with this difficulty and compare it with the aperture-scanning method. Although the experiment uses a model specimen with just two distinct layers separated along the optic axis, it should in principle be extendable to continuous objects.
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We reconsider the closed form solution of the ptychographic phase problem called the Wigner Distribution Deconvolution Method (WDDM), which has remained discarded for twenty years. Ptychographic reconstruction is nowadays always undertaken by iterative algorithms. WDDM gives rise to a 4 dimensional data cube of all the relative phases between points in the diffraction plane. Here we demonstrate a novel method to use all this information, instead of just the small subset used in the original 'stepping out' procedure developed in the 1990s, thus greatly suppressing noise. We further develop a method for designing an improved probe (illumination function) to further decrease noise effects during the deconvolution division. Combining these two with an iterative procedure for the deconvolution, which avoids the usual difficulty of a divide by a small number, we show in model calculations that WDDM competes well with the modern conventional iterative methods like ePIE (the extended Ptychographical Iterative Engine).
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We show for the first time that ptychography (a form of lensless diffractive imaging) can recover the spectral response of an object through simultaneous reconstruction of multiple images that represent the object's response to a particular mode present in the illumination. We solve the phase problem for each mode independently, even though the intensity arriving at every detector pixel is an incoherent superposition of several uncorrelated diffracted waves. Until recently, the addition of incoherent modes has been seen as a nuisance in diffractive imaging: here we show that not only can the difficulties they pose be removed, but that they can also be used to discover much more information about the object. If the illumination function is also mode-specific, we show that we can also solve simultaneously for a multiplicity of such illumination modes. The work opens exciting possibilities for information multiplexing in ptychography over all visible, X-ray and electron wavelengths.
RESUMEN
The ptychographical iterative engine (or PIE) is a recently developed phase retrieval algorithm that employs a series of diffraction patterns recorded as a known illumination function is translated to a set of overlapping positions relative to a target sample. The technique has been demonstrated successfully at optical and X-ray wavelengths and has been shown to be robust to detector noise and to converge considerably faster than support-based phase retrieval methods. In this paper, the PIE is extended so that the requirement for an accurate model of the illumination function is removed.