Azeem Ahmad

In this Letter, a high-fidelity single-shot differential quantitative phase microscopy (dQPM) method is presented to effectively image nearly transparent biological samples. The proposed method is based on a common-path Sagnac interferometric configuration, which provides superior temporal phase stability and robustness against environmental disturbances. The proposed system exploits a pseudothermal source to achieve high spatial sensitivity and generate dense interference fringes for effective single-shot differential quantitative phase imaging. The effectiveness of the proposed system is experimentally demonstrated with various samples, including silica microspheres, a USAF phase target, fixed and live HeLa cells, and mouse kidney tissue.

The online version contains supplementary material available at 10.1038/s41598-026-46064-1.

One representative of state-of-the-art full-field optical metrology techniques is fringe-pattern-based methods, where the measurand is encoded in the phase distribution of the recorded spatially-periodic intensity pattern. The phase demodulation process comprises a crucial part of a measurement defining the overall achieved accuracy, and its algorithm imposes one of the main information throughput limitations of the entire experimental unit. In this paper, we propose a novel solution that increases the phase space–bandwidth product (SBP) of the optical system in a purely numerical way. It is based on deep learning and a convolutional neural network called DeepQuadrature. It takes two images as the input data: the prefiltered fringe pattern (denoised and detrended by variational image decomposition, for example) and the local orientation map (modulo 2π). The network’s output is the quadrature function (input fringe pattern shifted in phase by π/2). The phase distribution is finally calculated via the arctangent function of input and output fringe patterns. The working principle is somewhat motivated by the state of the art in high SBP classical phase estimation, called Hilbert spiral transform, however, with novel adaptive spectral features promoting DeepQuadrature as a versatile and universally applicable method. Phase decoding results for widespread simulated and experimental (testing technical and biological objects) fringe data compare favorably with classical algorithms: learning-free variational Hilbert quantitative phase imaging and the reference multi-frame method. DeepQuadrature thus opens new possibilities in fringe-based high-content optical metrology and is poised to advance biomedical and technical label-free microscopy.

Extended depth of focus (DOF) with high lateral resolution is the primary requirement of the transducer in scanning acoustic microscopy to generate high-resolution images of the three-dimensional sample over a large depth. Traditionally, focused ultrasonic spherical transducers are used to tightly focus the acoustic waves generated from a piezoelectric material for a wide range of applications in industrial, medical, and other fields. Such transducers have a problem of narrow DOF which restricts the imaging range in depth. In the present work, we propose three different transducer designs such as single axicon, central flat axicon, and double axicon, which enable the possibilities of high transverse resolution imaging over greater depths due to the significant increase in DOF. Finite element modeling (FEM) in comsol of a spherical, single axicon, central flat axicon, and double axicon transducer is systematically performed and compared in terms of transverse resolution, DOF, and acoustic pressure in the central lobe. In addition, the single axicon and double axicon transducer modeling is done for different apex angles. It is observed that the central flat axicon transducer allows customizable DOF and the double axicon transducer provides high lateral resolution and reduced pressure in the side lobes compared to a single axicon lens.

This paper presents the development of a single-shot, partially spatially coherent quantitative differential phase contrast microscopy (Q-DPCM) system. The optical scheme comprises a polarizer, lenses, calcite beam displacer, and analyzer, which can be seamlessly integrated to an existing bright-field microscopy system, transforming it into a Q-DPCM system. It utilizes a partially spatially coherent light source, enabling single-shot quantitative differential phase recovery of the specimens with high spatial phase sensitivity. It generates highly sensitive quantitative differential phase images of the specimens along one direction, like a gradient light interference microscopy (GLIM) system, using only a single interferogram. First, we validated the differential phase measurement capability of the system through experiments on polystyrene spheres (diameter 5.2 µm) and HeLa cells. Next, the system is utilized to generate quantitative phase maps of human red blood cells using two orthogonal differential interferograms recorded at two orientations of the calcite beam displacer. Further, the Q-DPCM system is implemented for 1-h time-lapse live cell monitoring, revealing the dynamics of intracellular granules such as nucleolus and lipids in U2OS cells.

Current low coherence quantitative phase microscopy (LC-QPM) systems suffer from either reduced field of view (FoV) or reduced temporal resolution due to the short temporal coherence (TC) length of the light source. Here, we propose a hybrid, experimental and numerical approach to address this core problem associated with LC-QPM. We demonstrate high spatial resolution and high phase sensitivity in LC-QPM at high temporal resolution. High space–time bandwidth product is achieved by employing incoherent light source for sample illumination in QPM to increase the spatial resolution and single-shot Hilbert spiral transform (HST) based phase recovery algorithm to enhance the temporal resolution without sacrificing spatial resolution during the reconstruction steps. The high spatial phase sensitivity comes by default due to the use of incoherent light source in QPM which has low temporal coherence length and does not generate speckle noise and coherent noise. The spatial resolution achieved by the HST is slightly inferior to the temporal phase-shifting (TPS) method when tested on a specimen but surpasses that of the single-shot Fourier transform (FT) based phase recovery method. Contrary to HST method, FT method requires high density fringes for lossless phase recovery, which is difficult to achieve in LC-QPM over entire FoV. Consequently, integration of HST algorithm with LC-QPM system makes an attractive route. Here, we demonstrate scalable FoV and resolution in single-shot LC-QPM and experimentally corroborate it on a test object and on both live and fixed biological specimen such as MEF, U2OS and human red blood cells (RBCs). LC-QPM system with HST reconstruction offer high-speed single-shot QPM imaging at high phase sensitivity and high spatial resolution enabling us to study sub-cellular dynamic inside U2OS for extended duration (3 h) and observe high-speed (50 fps) dynamics of human RBCs. The experimental results validate the effectiveness of the present approach and will open new avenues in the domain of biomedical imaging in the future.

With applications ranging from metabolomics to histopathology, quantitative phase microscopy (QPM) is a powerful label-free imaging modality. Despite significant advances in fast multiplexed imaging sensors and deep-learning-based inverse solvers, the throughput of QPM is currently limited by the pixel-rate of the image sensors. Complementarily, to improve throughput further, here we propose to acquire images in a compressed form so that more information can be transferred beyond the existing hardware bottleneck of the image sensor. To this end, we present a numerical simulation of a learnable optical compression-decompression framework that learns content-specific features. The proposed differentiable quantitative phase microscopy (∂-QPM) first uses learnable optical processors as image compressors. The intensity representations produced by these optical processors are then captured by the imaging sensor. Finally, a reconstruction network running on a computer decompresses the QPM images post aquisition. In numerical experiments, the proposed system achieves compression of × 64 while maintaining the SSIM of ∼0.90 and PSNR of ∼30 dB on cells. The results demonstrated by our experiments open up a new pathway to QPM systems that may provide unprecedented throughput improvements.

This study aims to demonstrate the generation and detection of Scholte waves inside polystyrene microparticles. This was proven using both experimental analysis and COMSOL simulation. Microspheres of different sizes were excited optically with a pulsed laser (532 nm), and the acoustic signals were detected using a transducer (40 MHz). On analyzing the laser-generated ultrasound signals, the results obtained experimentally and from COMSOL are in close agreement both in the time and frequency domain. A simplified analysis of Scholte wave generation by laser irradiation for homogeneous, isotropic microspheres is presented. The theoretical wave velocity of the Scholte wave was calculated and found close to our experimental results. A representation of pressure wave motion showing the Scholte wave generation is presented at different times.

We report three-dimensional surface profilometry with extended range of height measurements using synthetic wavelength scanning interferometry without tunable filters, wavelength-tuning lasers, grating elements. We have used inexpensive multiple colour light emitting diodes (LEDs) and operate them sequentially one by one or combination of two or more colours simultaneously to visualize synthetic wavelength light source. Multiple colour LED light source was synthesized and entire visible range from violet to deep red colour was covered. A wide range of synthetic wavelengths were obtained. Five step phase shifting algorithm was used to recover phase maps with Mirau type interferometer which can be further utilized for 3D height measurements. A simple phase subtraction method was used to reconstruct the phase map at synthetic wavelength. The present system provides extended range of height measurements from sub-wavelength to tens of wavelengths. Experimental results of 3D-surface profile measurements of Si-IC chip and standard step object are presented.