Volume 29 Issue 10 | Journal of Biomedical Optics

Journal of Biomedical Optics

VOL. 29 · NO. 10 | October 2024

CONTENTS

IN THIS ISSUE

General (2)

Imaging (2)

Microscopy (2)

Sensing (1)

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General

Quantitative estimation of optical properties in bilayer media within the subdiffusive regime using a tilted fiber-optic probe in diffuse reflectance spectroscopy, part 1: a theoretical framework for designing probe geometry

Philippe De Tillieux, Maxime Baillot, Pierre Marquet

Journal of Biomedical Optics, Vol. 29, Issue 10, 105001, (October 2024) https://doi.org/10.1117/1.JBO.29.10.105001

TOPICS: Scattering, Tissues, Photons, Reflectivity, Optical properties, Diffuse reflectance spectroscopy, Inverse problems, Tissue optics, Inverse optics, Monte Carlo methods

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Quantitative estimation of optical properties in bilayer media within the subdiffusive regime using tilted fiber-optic probe diffuse reflectance spectroscopy, part 2: probe design, realization, and experimental validation

Philippe De Tillieux, Maxime Baillot, Pierre Marquet

Journal of Biomedical Optics, Vol. 29, Issue 10, 105002, (October 2024) https://doi.org/10.1117/1.JBO.29.10.105002

TOPICS: Error analysis, Optical properties, Photons, Simulations, Melanoma, Tissues, Reflectivity, Design, Light sources and illumination, Statistical analysis

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Imaging

Dynamic multispectral NIR/SWIR for in vivo lymphovascular architectural and functional quantification

Christopher Hansen, Jaidip Jagtap, Abdul Parchur, Gayatri Sharma, Shayan Shafiee, Sayantan Sinha, Heather Himburg, Amit Joshi

Journal of Biomedical Optics, Vol. 29, Issue 10, 106001, (September 2024) https://doi.org/10.1117/1.JBO.29.10.106001

TOPICS: Lymphatic system, Short wave infrared radiation, Biological imaging, In vivo imaging, Fluorescence imaging, Imaging systems, Image resolution, Quantum imaging, Silver, Spatial resolution

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Mueller matrix analysis of a biologically sourced engineered tissue construct as polarimetric phantom

Zixi Lin, Samantha Madnick, Joshua Burrow, Jeffrey Morgan, Kimani Toussaint Jr.

Journal of Biomedical Optics, Vol. 29, Issue 10, 106002, (October 2024) https://doi.org/10.1117/1.JBO.29.10.106002

TOPICS: Tissues, Polarimetry, Collagen, Depolarization, Mueller matrices, Second harmonic generation, Biological samples, Biological research, Polarization, Biological imaging

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Significance

The polarimetric properties of biological tissues are often difficult to ascertain independent of their complex structural and organizational features. Conventional polarimetric tissue phantoms have well-characterized optical properties but are overly simplified. We demonstrate that an innovative, biologically sourced, engineered tissue construct better recapitulates the desired structural and polarimetric properties of native collagenous tissues, with the added benefit of potential tunability of the polarimetric response. We bridge the gap between non-biological polarimetric phantoms and native tissues.

Aim

We aim to evaluate a synthesized tissue construct for its effectiveness as a phantom that mimics the polarimetric properties in typical collagenous tissues.

Approach

We use a fibroblast-derived, ring-shaped engineered tissue construct as an innovative tissue phantom for polarimetric imaging. We perform polarimetry measurements and subsequent analysis using the Mueller matrix decomposition and Mueller matrix transformation methods. Scalar polarimetric parameters of the engineered tissue are analyzed at different time points for both a control group and for those treated with the transforming growth factor (TGF)-β1. Second-harmonic generation (SHG) imaging and three-dimensional collagen fiber organization analysis are also applied.

Results

We identify linear retardance and circular depolarization as the parameters that are most sensitive to the tissue culture time and the addition of TGF-β1. Aside from a statistically significant increase over time, the behavior of linear retardance and circular depolarization indicates that the addition of TGF-β1 accelerates the growth of the engineered tissue, which is consistent with expectations. We also find through SHG images that collagen fiber organization becomes more aligned over time but is not susceptible to the addition of TGF-β1.

Conclusions

The engineered tissue construct exhibits changes in polarimetric properties, especially linear retardance and circular depolarization, over culture time and under TGF-β1 treatments. This tissue construct has the potential to act as a controlled modular optical phantom for polarimetric-based methods.

Microscopy

Simultaneous assessment of NAD(P)H and flavins with multispectral fluorescence lifetime imaging microscopy at a single excitation wavelength of 750 nm

Boris Yakimov, Anastasia Komarova, Elena Nikonova, Artem Mozherov, Liubov Shimolina, Marina Shirmanova, Wolfgang Becker, Evgeny Shirshin, Vladislav Shcheslavskiy

Journal of Biomedical Optics, Vol. 29, Issue 10, 106501, (September 2024) https://doi.org/10.1117/1.JBO.29.10.106501

TOPICS: Fluorescence, Fluorescence lifetime imaging, Nicotinamide adenine dinucleotide, Emission wavelengths, Mode conditioning cables, Sensors, Fluorescence intensity, Cancer, Microscopy, Tumors

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Significance

Autofluorescence characteristics of the reduced nicotinamide adenine dinucleotide and oxidized flavin cofactors are important for the evaluation of the metabolic status of the cells. The approaches that involve a detailed analysis of both spectral and time characteristics of the autofluorescence signals may provide additional insights into the biochemical processes in the cells and biological tissues and facilitate the transition of spectral fluorescence lifetime imaging into clinical applications.

Aim

We present the experiments on multispectral fluorescence lifetime imaging with a detailed analysis of the fluorescence decays and spectral profiles of the reduced nicotinamide adenine dinucleotide and oxidized flavin under a single excitation wavelength aimed at understanding whether the use of multispectral detection is helpful for metabolic imaging of cancer cells.

Approach

We use two-photon spectral fluorescence lifetime imaging microscopy. Starting from model solutions, we switched to cell cultures treated by metabolic inhibitors and then studied the metabolism of cells within tumor spheroids.

Results

The use of a multispectral detector in combination with an excitation at a single wavelength of 750 nm allows the identification of fluorescence signals from three components: free and bound NAD(P)H, and flavins based on the global fitting procedure. Multispectral data make it possible to assess not only the lifetime but also the spectral shifts of emission of flavins caused by chemical perturbations. Altogether, the informative parameters of the developed approach are the ratio of free and bound NAD(P)H amplitudes, the decay time of bound NAD(P)H, the amplitude of flavin fluorescence signal, the fluorescence decay time of flavins, and the spectral shift of the emission signal of flavins. Hence, with multispectral fluorescence lifetime imaging, we get five independent parameters, of which three are related to flavins.

Conclusions

The approach to probe the metabolic state of cells in culture and spheroids using excitation at a single wavelength of 750 nm and a fluorescence time-resolved spectral detection with the consequent global analysis of the data not only simplifies image acquisition protocol but also allows to disentangle the impacts of free and bound NAD(P)H, and flavin components evaluate changes in their fluorescence parameters (emission spectra and fluorescence lifetime) upon treating cells with metabolic inhibitors and sense metabolic heterogeneity within 3D tumor spheroids.

High-resolution lensless holographic microscopy using a physics-aware deep network

Ashwini S. Galande, Vikas Thapa, Aswathy Vijay, Renu John

Journal of Biomedical Optics, Vol. 29, Issue 10, 106502, (October 2024) https://doi.org/10.1117/1.JBO.29.10.106502

TOPICS: Holograms, Education and training, Holography, Microscopy, Phase reconstruction, Data modeling, Wave propagation, 3D image reconstruction, Physics, Biological samples

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Significance

Lensless digital inline holographic microscopy (LDIHM) is an emerging quantitative phase imaging modality that uses advanced computational methods for phase retrieval from the interference pattern. The existing end-to-end deep networks require a large training dataset with sufficient diversity to achieve high-fidelity hologram reconstruction. To mitigate this data requirement problem, physics-aware deep networks integrate the physics of holography in the loss function to reconstruct complex objects without needing prior training. However, the data fidelity term measures the data consistency with a single low-resolution hologram without any external regularization, which results in a low performance on complex biological data.

Aim

We aim to mitigate the challenges with trained and physics-aware untrained deep networks separately and combine the benefits of both methods for high-resolution phase recovery from a single low-resolution hologram in LDIHM.

Approach

We propose a hybrid deep framework (HDPhysNet) using a plug-and-play method that blends the benefits of trained and untrained deep models for phase recovery in LDIHM. The high-resolution phase is generated by a pre-trained high-definition generative adversarial network (HDGAN) from a single low-resolution hologram. The generated phase is then plugged into the loss function of a physics-aware untrained deep network to regulate the complex object reconstruction process.

Results

Simulation results show that the SSIM of the proposed method is increased by 0.07 over the trained and 0.04 over the untrained deep networks. The average phase-SNR is elevated by 8.2 dB over trained deep models and 9.8 dB over untrained deep networks on the experimental biological cells (cervical cells and red blood cells).

Conclusions

We showed improved performance of the HDPhysNet against the unknown perturbation in the imaging parameters such as the propagation distance, the wavelength of the illuminating source, and the imaging sample compared with the trained network (HDGAN). LDIHM, combined with HDPhysNet, is a portable and technology-driven microscopy best suited for point-of-care cytology applications.

Sensing

Accuracy enhancement of metabolic index-based blood glucose estimation with a screening process for low-quality data

Tomoya Nakazawa, Keiji Morishita, Anna Ienaka, Takeo Fujii, Masaki Ito, Fumie Matsushita

Journal of Biomedical Optics, Vol. 29, Issue 10, 107001, (October 2024) https://doi.org/10.1117/1.JBO.29.10.107001

TOPICS: Signal to noise ratio, Glucose, Near infrared spectroscopy, Blood, Prototyping, Error analysis, Education and training, Sensors, Data acquisition, Background noise

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