Single-shot slightly off-axis digital holographic microscopy with add-on module based on beamsplitter cube

Picazo-Bueno, José Ángel; Trusiak, Maciej; Micó, Vicente

doi:10.1364/oe.27.005655

Cited by 64 publications

(25 citation statements)

References 82 publications

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“…The π-SMIM adds high-speed (video framerate) accurate and robust QPI capability to a commercial nonholographic microscope with enhanced phase reconstruction by properly placing a beam-splitter to record in a single-shot two holograms mutually phase shifted by π radians. Although we have previously reported on a single-shot approach for QPI using a beam-splitter interferometric architecture, 32 the method here presented is conceptually different from the previous one in the following sense. The previous approach 32 was targeted to provide an external add-on module to be adapted into the exit port of a regular microscope in order to update it with coherence sensing capabilities.…”

Section: Introductionmentioning

confidence: 90%

“…Although we have previously reported on a single-shot approach for QPI using a beam-splitter interferometric architecture, 32 the method here presented is conceptually different from the previous one in the following sense. The previous approach 32 was targeted to provide an external add-on module to be adapted into the exit port of a regular microscope in order to update it with coherence sensing capabilities. This external module is based on a beam-splitter cube interferometer working in slightly off-axis configuration, but the approach 32 does not imply internal modifications of the microscope as one needs in SMIM.…”

Section: Introductionmentioning

confidence: 90%

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Single-shot two-frame π-shifted spatially multiplexed interference phase microscopy

et al. 2019

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Single-shot, two-frame, π-shifted spatially multiplexed interference microscopy (π-SMIM) is presented as an improvement to previous SMIM implementations, introducing a versatile, robust, fast, and accurate method for cumbersome, noisy, and low-contrast phase object analysis. The proposed π-SMIM equips a commercially available nonholographic microscope with a high-speed (video frame rate) enhanced quantitative phase imaging (QPI) capability by properly placing a beam-splitter in the microscope embodiment to simultaneously (in a single shot) record two holograms mutually phase shifted by π radians at the expense of reducing the field of view. Upon subsequent subtractive superimposition of holograms, a π-hologram is generated with reduced background and improved modulation of interference fringes. These features determine superior phase retrieval quality, obtained by employing the Hilbert spiral transform on the π-hologram, as compared with a single low-quality (low signal-to-noise ratio) hologram analysis. In addition, π-SMIM enables accurate in-vivo analysis of high dynamic range phase objects, otherwise measurable only in static regime using time-consuming phase-shifting. The technique has been validated utilizing a 20 × ∕0.46 NA objective in a regular Olympus BX-60 upright microscope for QPI of different lines of prostate cancer cells and flowing microbeads.

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Section: Introductionmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 90%

Single-shot two-frame π-shifted spatially multiplexed interference phase microscopy

et al. 2019

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“…37,45,46 In this paper, we propose the common-path off-axis configuration by employing a cube beam splitter, which divides the incoming object beam into two beams, and a pinhole is used to filter spatially one of the beams at its Fourier plane, serving a reference beam. The beam splitter-based common-path DHM systems have been reported previously, 44,47 where the system 44 is based on the self-referencing method, and in Ref. 47, two π phase-shifted holograms are recorded and these two holograms are processed by using an algorithm based on the subtraction of the two fields of view (FOVs) in order to eliminate the direct current (DC) term.…”

Section: Introductionmentioning

confidence: 99%

“…The beam splitter-based common-path DHM systems have been reported previously, 44,47 where the system 44 is based on the self-referencing method, and in Ref. 47, two π phase-shifted holograms are recorded and these two holograms are processed by using an algorithm based on the subtraction of the two fields of view (FOVs) in order to eliminate the direct current (DC) term. These approaches are facing two major problems: (1) there is a possibility of overlapping of the object information at the image plane and may lead to generating twin images of the object and (2) if the first problem is overcome by utilizing only one half of the incident beam for object visualization, and another half is used for the reference beam, then, this process reduces the overall FOVs of the system by half.…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, in Ref. 47, since two holograms are recorded simultaneously by utilizing the half of the sensor area for the transmitted object beam from the beam splitter and another half area by the reflected object beam, which is π phase-shifted by the transmitted beam, it reduces the FOV of the system. Therefore, these problems can be overcome by spatially filtering one of the beams at its Fourier plane using a pinhole, obtained after passing through the beam splitter.…”

Section: Introductionmentioning

confidence: 99%

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Common-path multimodal three-dimensional fluorescence and phase imaging system

et al. 2020

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A stable multimodal system is developed by combining two common-path digital holographic microscopes (DHMs): coherent and incoherent, for simultaneous recording and retrieval of three-dimensional (3-D) phase and 3-D fluorescence imaging (FI), respectively, of a biological specimen. The 3-D FI is realized by a single-shot common-path off-axis fluorescent DHM developed recently by our group. In addition, we accomplish, the phase imaging by another single-shot, highly stable common-path off-axis DHM based on a beam splitter. In this DHM configuration, a beam splitter is used to divide the incoming object beam into two beams. One beam serves as the object beam carrying the useful information of the object under study, whereas another beam is spatially filtered at its Fourier plane by using a pinhole and it serves as a reference beam. This DHM setup, owing to a common-path geometry, is less vibration-sensitive and compact, having a similar field of view but with high temporal phase stability in comparison to a two-beam Mach-Zehnder-type DHM. The performance of the proposed common-path DHM and the multimodal system is verified by conducting various experiments on fluorescent microspheres and fluorescent protein-labeled living cells of the moss Physcomitrella patens. Moreover, the potential capability of the proposed multimodal system for 3-D live fluorescence and phase imaging of the fluorescent beads is also demonstrated. The obtained experimental results corroborate the feasibility of the proposed multimodal system and indicate its potential applications for the analysis of functional and structural behaviors of a biological specimen and enhancement of the understanding of physiological mechanisms and various biological diseases. © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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