We propose a single-frame lensfree phase retrieval(SFLFPR) method based on coherence-limited light-emitting diode (LED) illumination. Combining multi-wavelength scanning iteration for broad-spectrum illumination with phase support constraint, SFLFPR corrects resolution loss caused by the temporal coherence of LED, obtaining quantitative phase imaging results. Using only one hologram, our method can retrieve the high signal-to noise(SNR) phase of the sample and achieve a half-width resolution of 977 nm across a large field-of-view (FOV) of 19.53 mm2 , surpassing 1.41 times the resolution achieved by the conventional single-frame method. We confirmed the effectiveness of this method in quantitative phase imaging (QPI) by measuring various label-free samples including polystyrene microspheres, phase resolution target (PRT) and HeLa cell slices. Considering its fast real-time single-frame imaging capability, our method has a wide range of biological and medical applications.
We report a novel wide-field lens-free 3D microscopy, which is based on Fourier ptychography diffraction tomography (FPDT) technique. This method uses only one illumination angle to obtain a large enough number of diffractograms by scanning a wide range of wavelengths, applying an iterative method to fill the 3D spectrum, and finally recovering the refractive index (RI) distribution of the sample. And the effectiveness of the method in the 3D RI distribution reconstruction of a tilted phase target is experimentally verified.
Transport of intensity equation (TIE) is a well-established non-interferometric phase retrieval approach that enables quantitative phase imaging (QPI) by simply measuring intensity images at multiple axially displaced planes. The advantage of a TIE-based QPI system is its compatibility with partially coherent illumination, which provides speckle-free imaging with resolution beyond the coherent diffraction limit. However, TIE is generally implemented with a brightfield (BF) configuration, and the maximum achievable imaging resolution is still limited to the incoherent diffraction limit (twice the coherent diffraction limit). It is desirable that TIE-related approaches can surpass this limit and achieve high-throughput [high-resolution and wide field of view (FOV)] QPI. We propose a hybrid BF and darkfield transport of intensity (HBDTI) approach for high-throughput quantitative phase microscopy. Two through-focus intensity stacks corresponding to BF and darkfield illuminations are acquired through a low-numerical-aperture (NA) objective lens. The high-resolution and large-FOV complex amplitude (both quantitative absorption and phase distributions) can then be synthesized based on an iterative phase retrieval algorithm taking the coherence model decomposition into account. The effectiveness of the proposed method is experimentally verified by the retrieval of the USAF resolution target and different types of biological cells. The experimental results demonstrate that the half-width imaging resolution can be improved from 1230 nm to 488 nm with 2.5 × expansion across a 4 × FOV of 7.19 mm2, corresponding to a 6.25 × increase in space-bandwidth product from ∼5 to ∼30.2 megapixels. In contrast to conventional TIE-based QPI methods where only BF illumination is used, the synthetic aperture process of HBDTI further incorporates darkfield illuminations to expand the accessible object frequency, thereby significantly extending the maximum available resolution from 2NA to ∼5NA with a ∼5 × promotion of the coherent diffraction limit. Given its capability for high-throughput QPI, the proposed HBDTI approach is expected to be adopted in biomedical fields, such as personalized genomics and cancer diagnostics.
Transport of intensity equation (TIE) is an established quantitative phase imaging (QPI) method as a deter-ministic phase retrieval algorithm derived from paraxial approximation. However, this approximation limits TIE to retrieve high-resolution QPI for most biological samples. It can be overcome via the phase transfer function (PTF) method (no paraxial approximation), while it has incorrect phase in low-frequency due to the weak phase approximation. For the general microscopic specimens with thickness and small details, we present a resolution-enhanced QPI mixed-transfer-function (MTF) approach based on the spatial domain and the spatial frequency one, using phase space theory as a bridge. It is proved that the traditional TIE can obtain the accurate phase in low-frequency, but suffers from attenuation and blurring at high frequencies. Thus, MTF is combined with PTF to improve the maximum resolution that can be accurately reconstructed. Simulation results and experiments demonstrate that MTF can achieve high-contrast and high-resolution QPI correctly over the whole theoretical bandwidth, showing efficiency for phase retrieval even in slowly varying large phase objects. Besides, it is compatible with commercial microscopes without additional hardware modification, offering a flexible and cost-effective alternative for biomedical research and cellular investigations.
Transport of intensity equation (TIE) is an established quantitative phase imaging (QPI) method derived from paraxial approximation, however, this approximation limits TIE to retrieve high-resolution QPI. Thus, we present a dark-field transport of intensity equation (DFTIE) approach to improve resolution, solving the attenuation and blurring at high frequencies, which is combined iterative TIE phase retrieval with high-angle illumination coherent-mode decomposition to improve the maximum resolution under low numerical aperture objective. Simulation results demonstrate that DFTIE can achieve high-contrast and high-resolution QPI over the whole theoretical bandwidth, showing efficiency for high-throughput imaging. With the simple mode (only bright-field and dark-field are required) and multiplexing illumination, the imaging signal-to-noise ratio is higher, offering a flexible and cost-effective alternative for biomedical research and cellular investigations.
In traditional lensless in-line holographic microscopy, phase recovery based on multi-defocus distance is a common pixel-super-resolution technique. This method usually requires accurate displacement as a predictive condition, and the introduction of a precision mechanical displacement platform can bring about accurate displacement, but makes a simple lensless system complicated and expensive. In this paper, the samples are illuminated by a nearly coherent illumination system, and holograms at different heights are captured by the sensor driven by a low-cost servo motor without feedback system. Inaccurate displacement interval results in poor phase recovery. We propose a multi-height phase recovery algorithm based on z-axis correction to recover the phase information of the sample, which can improve the result by 1.58 times compared with that before correction. The reconstruction USAF target demonstrates a half-pitch lateral resolution of 775 nm across a large field-ofview of ∼29.84 mm2 , surpassing 2.15 times that of the theoretical Nyquist–Shannon sampling resolution limit imposed by the pixel size of the imaging sensor (1.67 μm).
According to the phase gradient transfer function (PGTF) derived from the phase space theory, the phase recovery algorithm based on the transport of intensity equation (TIE) has the problem that the high-frequency phase is underestimated due to the coherence effect of the limited aperture system under partially coherent illumination. Therefore, based on the theory of PGTF and phase transfer function (PTF), a phase reconstruction algorithm named high-resolution synthetic spectrum (HSS) method combining the TIE and the PTF-based deconvolution is proposed. This technique broadens the application range and provides high contrast, high accuracy, and highresolution quantitative phase results with high robustness. The performances of this technology are demonstrated by simulation and experiments, showing efficient for phase retrieval in the near-Fresnel region. Such a highresolution method can offer a flexible and cost-effective alternative for biomedical research and cell analysis, providing new avenues to design powerful computational imaging systems
We report a multi-wavelength multiplexed setup and associated super-resolution reconstruction method in lensless microscopy, which can generate high-resolution reconstructions from undersampled raw measurements captured at multiple wavelengths. The reconstruction result of the Benchmark Quantitative Phase Microscopy Target (QPTTM) demonstrates the resolution enhancement quantitatively, which achieves a half-pitch lateral resolution of 691 nm across a large field of view (~29.85 mm2), surpassing 2.41 times of the theoretical NyquistShannon sampling resolution limit imposed by the pixel-size of the sensor (1.67 µm). Compared with other superresolution methods such as lateral or axial shift-based device and illumination source rotation design, wavelength multiplexed avoids the need for shifting/rotating mechanical components. This multi-wavelength multiplexed super-resolution method would benet the research and development of a more stable lensless microscopy system.
Phase unwrapping is an essential procedure in digital holographic microscopy (DHM). There are many algorithms have been proposed to unwrap the phase such as the reliability-guided phase unwrapping algorithm that intro- duced in this paper. It is necessary to do a comparison of these algorithms in order to determine which method has better performance in the actual experiment. For higher quality and fewer error points, we also introduce an improved phase unwrapping path base on path-following method such as the reliability-guided phase unwrapping algorithm, and the experimental images demonstrate the validity of our algorithms. In addition, we propose a method to accelerate the phase unwrapping process for biomedical dynamic imaging. The experimental results suggest that this method can significantly improve the dynamic measurement speed while ensuring the accuracy of phase unwrapping.
We demonstrate a method for increasing the effective resolution of phase retrieval based on the transport of intensity equation (TIE) named speckle high-resolution synthetic spectrum (speckle-HSS), as the upgraded version of the speckle-TIE approach we proposed before based on the quantitative phase imaging camera with a weak diffuser (QPICWD). Benefit from the phase gradient transfer function (PGTF) and phase transfer function (PTF), the phase blurring caused by the underestimation of phase gradient can be compensated correctly via combining TIE and PTF-based deconvolution. This method broadens the application range, alleviating the artifacts and enhancing the contrast and resolution in more accurate value. The experimental results of live HeLa cells have been presented, showing the effectiveness of the proposed method.
In this paper, we present a multi-wavelength multiplexed setup and associated super-resolution reconstruction method in lensfree microscopy that generates high-resolution reconstructions from undersampled raw measurements captured at multiple wavelengths. The reconstruction result of the standard 1951 USAF achieves a half-pitch lateral resolution of 775 nm, corresponding to a numerical aperture of 1.0, across a large field of view (∼ 29.85 mm2). Compared with other super-resolution methods such as lateral or axial shift-based device and illumination source rotation design, wavelength multiplexed avoids the need for shifting/rotating mechanical components. This multi-wavelength multiplexed super-resolution method would benefit the research and development of a more stable lensfree microscopy system.
We present a wavelength-scanning-based lensfree microscopy that generates high-resolution reconstructions from undersampled raw measurements captured at multiple wavelengths.The reconstruction result of the standard 1951 USAF achieves a half-pitch lateral resolution of 775 nm, corresponding to a numerical aperture of ∼ 1.0, across a large field of view (∼ 29.85 mm2). Compared with other super-resolution methods such as lateral or axial shift-based device and illumination source rotation design, wavelength scanning avoids the need for shifting/rotating mechanical components. This wavelength-scanning super-resolution method would benefit the research and development of more stable lensfree microscopy system.
In this paper, a holographic lensless quantitative phase imaging (QPI) microscope is presented, which is composed of a CMOS detector image sensor with a programmable color LED matrix, without any lens and mechanical displacement device. Such a miniaturized system can provide a field-portable cost-effective platform for highthroughput quantification of multiple samples. Coordinating the self-developed software operating system, the bright-field imaging, the quantitative phase imaging as well as cell counting, profile analysis, three-dimensional (3D) imaging and differential interference contrast (DIC) imaging can be realized. With its high-resolution based computational microscopy interface, this system can be also adaptively used for telemedicine applications and point-of-care testing (POCT) in resource-limited environments.
Lensless imaging technique, as a newly developed microscopic imaging method, combined with its corresponding image restoration algorithm, and it can obtain large-field, high-resolution three-dimensional images without labeling. Therefore, the lensless imaging system has the advantages of low cost, good portability, large field of view and high resolution. However, due to the limitation of pixel size, hardware implementation and post data processing, the imaging resolution of lensless microscopy is far away from theoretical performance. In order to solve above problems, this paper proposes a method to improve the capability registration and information coupling based on multi-wavelength illumination. Moreover, this work combines a stack of captured low-resolution images into a high-resolution result image, and the pixel super-resolution can be realized throught the reconstruction algorithm based on the exist illumination light source, Finally, experimental results utilizing USAF target demonstrate the success of proposed lensless imaging method.
We present a novel approach to compensate coherence effect via combining the transport of intensity equation (TIE) with look-up table phase compensation (LUT-PC) method. It is the better version of the Speckle-TIE method we demonstrated before on the basis of the quantitative phase imaging camera with a weak diffuser (QPICWD). With the phase gradient ratio theory and the look-up table method, the phase blurring caused by underestimation of phase gradient will be compensated correctly by reasonable rescaling. The LUT-PC SpeckleTIE method has the evident predominance of speediness since it only needs one slightly defocused speckle image in one time owing to that the reference speckle image can be captured beforehand. The deblurring achieved by this method improves the imaging resolution to the theoretical partial coherence limit with good robustness, reducing artifacts and improving the accuracy and contrast. The experimental results show the effectiveness of the technique.
We present a new quantitative phase imaging method on the basis of the novel camera named quantitative phase imaging camera with a weak diffuser (QPICWD). It measures object under low-coherence quasi-monochromatic illumination via examining the deformation of the speckle intensity pattern. The speckle deformation can be analyzed by means of ensemble average of geometric flow method, realizing high resolution distortion field by using the transport of intensity equation (TIE). There are some applications for the proposed new design including nondestructive optical testing of microlens array with nanometric thickness. Since the proposed QPICWD needs no modification of the common bright-field microscope, it may promote QPI as a useful tool for subcellular level biological analysis.
We present an efficient quantitative phase imaging camera with a weak diffuser (QPICWD) based on the transport of intensity equation (TIE). The compact QPICWD measures object induced phase delay under low-coherence quasi-monochromatic illumination via examining the deformation of the speckle intensity pattern. Analysing the speckle deformation with an ensemble average of the geometric ow, we can achieve the high-resolution distortion field by the TIE. We present some applications for the proposed design involving nondestructive optical testing of microlens array with nanometric thickness and imaging of fixed and live unstained HeLa cells. Since the designed QPICWD needs no modification of the common bright-field microscope or additional accessories, it may advance QPI as a widely useful tool for biological analysis at subcellular levels.
In this paper, we employ coded aperture imaging (CAI), an emerging computational technology that captures 4D light-field information to realize pixel super-resolution imaging via post-processing. Our CAI experimental setup is built based on 4f delay system with reflective optical path structure, where a programmable LCOS spatial light modulator is integrated at the Fourier plane to implement high-resolution high-contrast aperture coding, without requiring specialized hardware or any moving parts. In addition, we propose an iterative super-solution reconstruction algorithm based on aperture coding, optical fields manipulation and compressed sensing. First, we establish an accurate mathematical model for the OTF of coded aperture system and pixel binning process. Then, based on a series of low-resolution intensity image, we computationally reconstruct the high-resolution image with the convex projection iterative algorithm. The effectiveness of this algorithm is demonstrated with both simulation and experimental results. Due to its flexibility and simplicity, this technology can break physical limitations of the detectors’ resolution to one that is solvable through computation, rendering it a promising tool in public security, military survey, medical science and many other fields.
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