Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells

Nygate, Yoav; Singh, Gyanendra; Barnea, Itay; Shaked, Natan T.

doi:10.1364/ol.43.002587

Cited by 23 publications

(14 citation statements)

References 11 publications

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“…In optical multiplexing [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30], multiple sample and reference beam pairs with different 𝜃 𝑥 and 𝜃 𝑦 combinations are projected onto the digital camera simultaneously, each of which creating an off-axis hologram with a different interference fringe direction that positions one wavefront in the SFD without overlapping other terms. The simultaneous projection of all beams on the camera may create unwanted interference between nonmatching pairs.…”

Section: Optical Multiplexingmentioning

confidence: 99%

Is multiplexed off-axis holography for quantitative phase imaging more spatial bandwidth-efficient than on-axis holography? [Invited]

Dardikman

Shaked

2018

J. Opt. Soc. Am. A

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Digital holographic microcopy is a thriving imaging modality that attracts considerable research interest due to its ability to not only create excellent label-free contrast, but also supply valuable physical information regarding the density and dimensions of the sample with nanometer-scale axial sensitivity. Three basic holographic recording geometries currently exist, including on-axis, off-axis and slightly off-axis holography, each of them enabling a variety of architectures in terms of bandwidth use and compression capacity. Specifically, off-axis holography and slightly off-axis holography allow spatial hologram multiplexing, enabling compressing more information into the same digital hologram. In this paper, we define an efficiency score used to analyze the various possible architectures, and compare the signal-to-noise ratio and mean squared error obtained using each of them, determining the optimal holographic method.

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Section: Optical Multiplexingmentioning

confidence: 99%

Is multiplexed off-axis holography for quantitative phase imaging more spatial bandwidth-efficient than on-axis holography? [Invited]

Dardikman

Shaked

2018

J. Opt. Soc. Am. A

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“…Various multimodal imaging systems with different configurations and for different purposes have been recently developed because these systems enable one to analyze the functional and structural behaviors of a biological specimen at a single examination and, therefore, facilitate a better understanding of the behavior of molecular, cellular, and disease biology. [1][2][3][4][5][6][7][8][9] The multimodal systems were developed for simultaneous fluorescence and quantitative phase imaging by incorporating the two-dimensional (2-D) epifluorescence microscopy with the diffraction phase microscopy by Park et al 1 and with the Mach-Zehnder-type digital holographic microscopy (DHM) by Pavillon et al 2 and Quan et al 3 A multimodal approach that incorporates confocal Raman, confocal reflectance, and quantitative phase microscopy 4 has demonstrated the potential for retrieving the molecular specific and morphological information. Optical diffraction tomography (ODT) is another technique to reconstruct the three-dimensional (3-D) quantitative phase imaging from the recorded multiple 2-D holograms.…”

Section: Introductionmentioning

confidence: 99%

“…The structured illumination microscopy-based multimodal system 7 is demonstrated for 3-D subdiffraction multimodal imaging of both quantitative phase and fluorescence. Chowdhury et al 8 and Nygate et al 9 proposed the multimodal systems based on the principle of off-axis holographic multiplexing to obtain the quantitative phase and fluorescence imaging (FI) of the biological cells using a single camera. These systems, 8,9 owing to the use of single camera for recording both the phase hologram and the fluorescence image, are free from image registration issues.…”

Section: Introductionmentioning

confidence: 99%

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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“…These techniques have been also integrated with deep learning to improve the space-bandwidth, automated classification of human red blood cells (RBC), spermatozoa, anthrax spores and among others [14][15][16][17]. However, the reconstruction algorithm associated with aforementioned techniques suffers with pixel limited resolution [11], twin image problem [12,13] and poor spatial phase sensitivity [18] thus cannot offer the fine structural information over the large field of view (FOV) of the specimens. Off-axis incoherent QPI i.e.…”

Section: Introductionmentioning

confidence: 99%

High space-bandwidth in quantitative phase imaging using partially spatially coherent digital holographic microscopy and a deep neural network

Butola¹,

Kanade²,

Bhatt³

et al. 2020

Opt. Express

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Quantitative phase microscopy (QPM) is a label-free technique that enables monitoring of morphological changes at the 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 quantitative phase imaging using partially spatially coherent digital holographic microscopy (PSC-DHM) assisted with a 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 the PSC-DHM 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 heterogeneous macrophages. The performance is evaluated by predicting the HR images from the datasets captured with a low NA lens and compared with the actual HR phase images. An improvement of 9× in the space-bandwidth product is demonstrated for both RBC and macrophages datasets. We believe that the PSC-DHM + 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.

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Simultaneous off-axis multiplexed holography and regular fluorescence microscopy of biological cells

Cited by 23 publications

References 11 publications

Is multiplexed off-axis holography for quantitative phase imaging more spatial bandwidth-efficient than on-axis holography? [Invited]

Is multiplexed off-axis holography for quantitative phase imaging more spatial bandwidth-efficient than on-axis holography? [Invited]

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

High space-bandwidth in quantitative phase imaging using partially spatially coherent digital holographic microscopy and a deep neural network

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