Xiaoze Ou

Quantitative phase imaging via Fourier ptychographic microscopy

Fourier ptychographic microscopy (FPM) is a recently developed imaging modality that uses angularly varying illumination to extend a systems performance beyond the limit defined by its optical components. The FPM technique applies a novel phase-retrieval procedure to achieve resolution enhancement and complex image recovery. In this Letter, we compare FPM data to theoretical prediction and phase-shifting digital holography measurement to show that its acquired phase maps are quantitative and artifact-free. We additionally explore the relationship between the achievable spatial and optical thickness resolution offered by a reconstructed FPM phase image. We conclude by demonstrating enhanced visualization and the collection of otherwise unobservable sample information using FPMs quantitative phase.

Embedded pupil function recovery for Fourier ptychographic microscopy

We develop and test a pupil function determination algorithm, termed embedded pupil function recovery (EPRY), which can be incorporated into the Fourier ptychographic microscopy (FPM) algorithm and recover both the Fourier spectrum of sample and the pupil function of imaging system simultaneously. This EPRY-FPM algorithm eliminates the requirement of the previous FPM algorithm for a priori knowledge of the aberration in the imaging system to reconstruct a high quality image. We experimentally demonstrate the effectiveness of this algorithm by reconstructing high resolution, large field-of-view images of biological samples. We also illustrate that the pupil function we retrieve can be used to study the spatially varying aberration of a large field-of-view imaging system. We believe that this algorithm adds more flexibility to FPM and can be a powerful tool for the characterization of an imaging systems aberration.

Aperture-scanning Fourier ptychography for 3D refocusing and super-resolution macroscopic imaging

We report an imaging scheme, termed aperture-scanning Fourier ptychography, for 3D refocusing and super-resolution macroscopic imaging. The reported scheme scans an aperture at the Fourier plane of an optical system and acquires the corresponding intensity images of the object. The acquired images are then synthesized in the frequency domain to recover a high-resolution complex sample wavefront; no phase information is needed in the recovery process. We demonstrate two applications of the reported scheme. In the first example, we use an aperture-scanning Fourier ptychography platform to recover the complex hologram of extended objects. The recovered hologram is then digitally propagated into different planes along the optical axis to examine the 3D structure of the object. We also demonstrate a reconstruction resolution better than the detector pixel limit (i.e., pixel super-resolution). In the second example, we develop a camera-scanning Fourier ptychography platform for super-resolution macroscopic imaging. By simply scanning the camera over different positions, we bypass the diffraction limit of the photographic lens and recover a super-resolution image of an object placed at the far field. This platforms maximum achievable resolution is ultimately determined by the cameras traveling range, not the aperture size of the lens. The FP scheme reported in this work may find applications in 3D object tracking, synthetic aperture imaging, remote sensing, and optical/electron/X-ray microscopy.

High numerical aperture Fourier ptychography: principle, implementation and characterization

Fourier ptychography (FP) utilizes illumination control and computational post-processing to increase the resolution of bright-field microscopes. In effect, FP extends the fixed numerical aperture (NA) of an objective lens to form a larger synthetic system NA. Here, we build an FP microscope (FPM) using a 40X 0.75NA objective lens to synthesize a system NA of 1.45. This system achieved a two-slit resolution of 335 nm at a wavelength of 632 nm. This resolution closely adheres to theoretical prediction and is comparable to the measured resolution (315 nm) associated with a standard, commercially available 1.25 NA oil immersion microscope. Our work indicates that Fourier ptychography is an attractive method to improve the resolution-versus-NA performance, increase the working distance, and enlarge the field-of-view of high-resolution bright-field microscopes by employing lower NA objectives.

Solving ptychography with a convex relaxation

Ptychography is a powerful computational imaging technique that transforms a collection of low-resolution images into a high-resolution sample reconstruction. Unfortunately, algorithms that currently solve this reconstruction problem lack stability, robustness, and theoretical guarantees. Recently, convex optimization algorithms have improved the accuracy and reliability of several related reconstruction efforts. This paper proposes a convex formulation of the ptychography problem. This formulation has no local minima, it can be solved using a wide range of algorithms, it can incorporate appropriate noise models, and it can include multiple a priori constraints. The paper considers a specific algorithm, based on low-rank factorization, whose runtime and memory usage are near-linear in the size of the output image. Experiments demonstrate that this approach offers a 25% lower background variance on average than alternating projections, the ptychographic reconstruction algorithm that is currently in widespread use.

Characterization of spatially varying aberrations for wide field-of-view microscopy

We describe a simple and robust approach for characterizing the spatially varying pupil aberrations of microscopy systems. In our demonstration with a standard microscope, we derive the location-dependent pupil transfer functions by first capturing multiple intensity images at different defocus settings. Next, a generalized pattern search algorithm is applied to recover the complex pupil functions at ~350 different spatial locations over the entire field-of-view. Parameter fitting transforms these pupil functions into accurate 2D aberration maps. We further demonstrate how these aberration maps can be applied in a phase-retrieval based microscopy setup to compensate for spatially varying aberrations and to achieve diffraction-limited performance over the entire field-of-view. We believe that this easy-to-use spatially-varying pupil characterization method may facilitate new optical imaging strategies for a variety of wide field-of-view imaging platforms.

Diffraction tomography with Fourier ptychography

This paper presents a technique to image the complex index of refraction of a sample across three dimensions. The only required hardware is a standard microscope and an array of LEDs. The method, termed Fourier ptychographic tomography (FPT), first captures a sequence of intensity-only images of a sample under angularly varying illumination. Then, using principles from ptychography and diffraction tomography, it computationally solves for the sample structure in three dimensions. The experimental microscope demonstrates a lateral spatial resolution of 0.39 μm and an axial resolution of 3.7 μm at the Nyquist-Shannon sampling limit (0.54 and 5.0 μm at the Sparrow limit, respectively) across a total imaging depth of 110 μm. Unlike competing methods, this technique quantitatively measures the volumetric refractive index of primarily transparent and contiguous sample features without the need for interferometry or any moving parts. Wide field-of-view reconstructions of thick biological specimens suggest potential applications in pathology and developmental biology.

Fourier ptychographic microscopy for filtration-based circulating tumor cell enumeration and analysis

Abstract. Circulating tumor cells (CTCs) are recognized as a candidate biomarker with strong prognostic and predictive potential in metastatic disease. Filtration-based enrichment technologies have been used for CTC characterization, and our group has previously developed a membrane microfilter device that demonstrates efficacy in model systems and clinical blood samples. However, uneven filtration surfaces make the use of standard microscopic techniques a difficult task, limiting the performance of automated imaging using commercially available technologies. Here, we report the use of Fourier ptychographic microscopy (FPM) to tackle this challenge. Employing this method, we were able to obtain high-resolution color images, including amplitude and phase, of the microfilter samples over large areas. FPM’s ability to perform digital refocusing on complex images is particularly useful in this setting as, in contrast to other imaging platforms, we can focus samples on multiple focal planes within the same frame despite surface unevenness. In model systems, FPM demonstrates high image quality, efficiency, and consistency in detection of tumor cells when comparing corresponding microfilter samples to standard microscopy with high correlation (R2=0.99932). Based on these results, we believe that FPM will have important implications for improved, high throughput, filtration-based CTC analysis, and, more generally, image analysis of uneven surfaces.

0.5 gigapixel microscopy using a flatbed scanner

The capability to perform high-resolution, wide field-of-view (FOV) microscopy imaging is highly sought after in biomedical applications. In this paper, we report a wide FOV microscopy system that uses a closed-circuit-television (CCTV) lens for image relay and a flatbed scanner for data acquisition. We show that such an imaging system is capable of capturing a 10 mm × 7.5 mm FOV image with 0.78 µm resolution, resulting in more than 0.5 billion pixels across the entire image. The resolution and field curve of the proposed system were characterized by imaging a USAF resolution target and a hole-array target. To demonstrate its application, 0.5 gigapixel images of histology slides were acquired using this system.

Overlapped Fourier coding for optical aberration removal.

We present an imaging procedure that simultaneously optimizes a cameras resolution and retrieves a samples phase over a sequence of snapshots. The technique, termed overlapped Fourier coding (OFC), first digitally pans a small aperture across a cameras pupil plane with a spatial light modulator. At each aperture location, a unique image is acquired. The OFC algorithm then fuses these low-resolution images into a full-resolution estimate of the complex optical field incident upon the detector. Simultaneously, the algorithm utilizes redundancies within the acquired dataset to computationally estimate and remove unknown optical aberrations and system misalignments via simulated annealing. The result is an imaging system that can computationally overcome its optical imperfections to offer enhanced resolution, at the expense of taking multiple snapshots over time.