Influence of laser spot size at diffuser plane on the longitudinal spatial coherence function of optical coherence microscopy system

Coherence properties and wavelength of light sources are indispensable for optical coherence microscopy/tomography as they greatly influence the signal to noise ratio, axial resolution, and penetration depth of the system. In the present letter, we investigated the longitudinal spatial coherence properties of the pseudo-thermal light source (PTS) as a function of spot size at the diffuser plane, which is controlled by translating microscope objective lens towards or away from the diffuser plane. The axial resolution of PTS is found to be maximum ~ 13 microns for the beam spot size of 3.5 mm at the diffuser plane. The change in the axial resolution of the system as the spot size is increased at the diffuser plane is further confirmed by performing experiments on standard gauge blocks of height difference of 15 microns. Thus, by appropriately choosing the beam spot size at the diffuser plane, any monochromatic laser light source depending on the biological window can be utilized to obtain high axial-resolution with large penetration depth and speckle-free tomographic images of multilayered biological specimens irrespective of the source temporal coherence length. In addition, PTS could be an attractive alternative light source for achieving high axial-resolution without needing chromatic aberration corrected optics and dispersion-compensation mechanism, unlike conventional setups.Comment: 11 pages, 4 figures. arXiv admin note: text overlap with arXiv:1810.0199

Characterization of color cross-talk of CCD detectors and its influence in multispectral quantitative phase imaging

Multi-spectral quantitative phase imaging (QPI) is an emerging imaging modality for wavelength dependent studies of several biological and industrial specimens. Simultaneous multi-spectral QPI is generally performed with color CCD cameras. However, color CCD cameras are suffered from the color crosstalk issue, which needed to be explored. Here, we present a new approach for accurately measuring the color crosstalk of 2D area detectors, without needing prior information about camera specifications. Color crosstalk of two different cameras commonly used in QPI, single chip CCD (1-CCD) and three chip CCD (3-CCD), is systematically studied and compared using compact interference microscopy. The influence of color crosstalk on the fringe width and the visibility of the monochromatic constituents corresponding to three color channels of white light interferogram are studied both through simulations and experiments. It is observed that presence of color crosstalk changes the fringe width and visibility over the imaging field of view. This leads to an unwanted non-uniform background error in the multi-spectral phase imaging of the specimens. It is demonstrated that the color crosstalk of the detector is the key limiting factor for phase measurement accuracy of simultaneous multi-spectral QPI systems.Comment: 16 pages, 8 figure

Effect on the longitudinal coherence properties of a pseudothermal light source as a function of source size and temporal coherence

© 2019 Optical Society of America. Users may use, reuse, and build upon the article, or use the article for text or data mining, so long as such uses are for non-commercial purposes and appropriate attribution is maintained. All other rights are reserved.In the present Letter, a synthesized pseudothermal light source having high temporal coherence (TC) and low spatial coherence (SC) properties is used. The longitudinal coherence (LC) properties of the spatially extended monochromatic light source are systematically studied. The pseudothermal light source is generated from two different monochromatic laser sources: He–Ne (at 632 nm) and DPSS (at 532 nm). It was found that the LC length of such a light source becomes independent of the parent laser’s TC length for a large source size. For the chosen lasers, the LC length becomes constant to about 30 μm for a laser source size of ≥3.3  mm ≥3.3  mm . Thus, by appropriately choosing the source size, any monochromatic laser light source depending on the biological window can be utilized to obtain high axial resolution in an optical coherence tomography (OCT) system irrespective of its TC length. The axial resolution of 650 nm was obtained using a 1.2 numerical aperture objective lens at a 632 nm wavelength. These findings pave the path for widespread penetration of pseudothermal light into existing OCT systems with enhanced performance. A pseudothermal light source with high TC and low SC properties could be an attractive alternative light source for achieving high axial resolution without needing dispersion compensation as compared to a broadband light source

High-throughput spatial sensitive quantitative phase microscopy using low spatial and high temporal coherent illumination

High space-bandwidth product with high spatial phase sensitivity is indispensable for a single-shot quantitative phase microscopy (QPM) system. It opens avenue for widespread applications of QPM in the field of biomedical imaging. Temporally low coherence light sources are implemented to achieve high spatial phase sensitivity in QPM at the cost of either reduced temporal resolution or smaller field of view (FOV). In addition, such light sources have low photon degeneracy. On the contrary, high temporal coherence light sources like lasers are capable of exploiting the full FOV of the QPM systems at the expense of less spatial phase sensitivity. In the present work, we demonstrated that use of narrowband partially spatially coherent light source also called pseudo-thermal light source (PTLS) in QPM overcomes the limitations of conventional light sources. The performance of PTLS is compared with conventional light sources in terms of space bandwidth product, phase sensitivity and optical imaging quality. The capabilities of PTLS are demonstrated on both amplitude (USAF resolution chart) and phase (thin optical waveguide, height ~ 8 nm) objects. The spatial phase sensitivity of QPM using PTLS is measured to be equivalent to that for white light source and supports the FOV (18 times more) equivalent to that of laser light source. The high-speed capabilities of PTLS based QPM is demonstrated by imaging live sperm cells that is limited by the camera speed and large FOV is demonstrated by imaging histopathology human placenta tissue samples. Minimal invasive, high-throughput, spatially sensitive and single-shot QPM based on PTLS will enable wider penetration of QPM in life sciences and clinical applications

Label-free imaging on waveguide platform with enhanced resolution and contrast

Chip-based Evanescent Light Scattering (cELS) utilizes the multiple modes of a high-index contrast optical waveguide for near-field illumination of unlabeled samples, thereby repositioning the highest spatial frequencies of the sample into the far-field. The multiple modes scattering off the sample with different phase differences is engineered to have random spatial distributions within the integration time of the camera, mitigating the coherent speckle noise. This enables label-free superior-contrast imaging of weakly scattering nanosized specimens such as extra-cellular vesicles (EVs) and liposomes, dynamics of living HeLa cells etc. We demonstrate a multi-moded straight waveguide as a partially coherent light source. For isotropic super-resolution, spatially incoherent light engineered via multiple-arms waveguide chip and intensity-fluctuation based algorithms are used. The proof-of-concept results are demonstrated on 100 nm polystyrene beads and resolution improvement of close to 2× is shown. cELS also realizes (2-10)× more contrast as opposed to conventional imaging techniques

High space-bandwidth in quantitative phase imaging using partially spatially coherent optical coherence microscopy and deep neural network

Quantitative phase microscopy (QPM) is a label-free technique that enables to monitor morphological changes at subcellular level. The performance of the QPM system in terms of spatial sensitivity and resolution depends on the coherence properties of the light source and the numerical aperture (NA) of objective lenses. Here, we propose high space-bandwidth QPM using partially spatially coherent optical coherence microscopy (PSC-OCM) assisted with deep neural network. The PSC source synthesized to improve the spatial sensitivity of the reconstructed phase map from the interferometric images. Further, compatible generative adversarial network (GAN) is used and trained with paired low-resolution (LR) and high-resolution (HR) datasets acquired from PSC-OCM system. The training of the network is performed on two different types of samples i.e. mostly homogenous human red blood cells (RBC) and on highly heterogenous macrophages. The performance is evaluated by predicting the HR images from the datasets captured with low NA lens and compared with the actual HR phase images. An improvement of 9 times in space-bandwidth product is demonstrated for both RBC and macrophages datasets. We believe that the PSC-OCM+GAN approach would be applicable in single-shot label free tissue imaging, disease classification and other high-resolution tomography applications by utilizing the longitudinal spatial coherence properties of the light source

Synthetic wavelength scanning interferometry for 3D surface profilometry with extended range of height measurement using multi-colour LED light sources

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