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Miniature, multi-dichroic instrument for measuring the concentration of multiple fluorophores 用于测量多种荧光团浓度的微型多分色仪器
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-14 DOI: 10.1364/boe.516574
Konstantin Grygoryev, Huihui Lu, Simon Sørensen, Omid Talebi Varnosfaderani, Rachel Georgel, Liyao Li, Ray Burke, and Stefan Andersson-Engels
Identification of tumour margins during resection of the brain is critical for improving the post-operative outcomes. Due to the highly infiltrative nature of glioblastoma multiforme (GBM) and limited intraoperative visualization of the tumour margin, incomplete surgical resection has been observed to occur in up to 80 % of GBM cases, leading to nearly universal tumour recurrence and overall poor prognosis of 14.6 months median survival. This research presents a miniaturized, SiPMT-based optical system for simultaneous measurement of powerful DRS and weak auto-fluorescence for brain tumour detection. The miniaturisation of the optical elements confined the spatial separation of eight select wavelengths into footprint measuring 1.5 × 2 × 16 mm. The small footprint enables this technology to be integrated with existing surgical guidance instruments in the operating room. It’s dynamic ability to subtract any background illumination and measure signal intensities across a broad range from pW to mWs make this design much more suitable for clinical environments as compared to spectrometer-based systems with limited dynamic ranges and high integration times. Measurements using optical tissue phantoms containing mixed fluorophores demonstrate correlation coefficients between the fitted response and actual concentration using PLS regression being 0.95, 0.87 and 0.97 for NADH, FAD and PpIX , respectively. These promising results indicate that our proposed miniaturized instrument could serve as an effective alternative in operating rooms, assisting surgeons in identifying brain tumours to achieving positive surgical outcomes for patients.
在脑部切除术中确定肿瘤边缘对改善术后效果至关重要。由于多形性胶质母细胞瘤(GBM)具有高度浸润性,而术中对肿瘤边缘的观察有限,据观察,高达 80% 的 GBM 病例会出现手术切除不彻底的情况,导致几乎普遍的肿瘤复发和中位生存期仅为 14.6 个月的不良预后。这项研究提出了一种基于 SiPMT 的微型光学系统,可同时测量用于脑肿瘤检测的强 DRS 和弱自发荧光。光学元件的微型化将八个选择波长的空间分离限制在 1.5 × 2 × 16 毫米的尺寸内。由于体积小,这项技术可以与手术室现有的手术引导仪器集成。与动态范围有限和积分时间较长的光谱仪系统相比,该系统能够动态地减去任何背景光照,并在从 pW 到 mWs 的宽范围内测量信号强度,因此更适合临床环境。使用含有混合荧光团的光学组织模型进行的测量表明,使用 PLS 回归法,NADH、FAD 和 PpIX 的拟合响应与实际浓度之间的相关系数分别为 0.95、0.87 和 0.97。这些令人鼓舞的结果表明,我们提出的微型仪器可作为手术室的有效替代品,协助外科医生识别脑肿瘤,为患者带来积极的手术效果。
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引用次数: 0
User-friendly oblique plane microscopy on a fully functional commercially available microscope base 在功能齐全的商用显微镜底座上使用方便的斜面显微镜
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-14 DOI: 10.1364/boe.518856
George Sirinakis, Edward S. Allgeyer, Dmitry Nashchekin, and Daniel St. Johnston
In this work we present an oblique plane microscope designed to work seamlessly with a commercially available microscope base. To support all the functionality offered by the microscope base, where the position of the objective lens is not fixed, we adopted a two-mirror scanning geometry that can compensate for changes to the position of the objective lens during routine microscope operation. We showed that within a ± 1 mm displacement range of the 100X, 1.35 NA objective lens away from its designed position, the PSF size increased by <3% and <11% in the lateral and axial dimensions, respectively, while the error in magnification was <0.5% within volumes extending ± 10 µm about the focal plane. Compared to the more traditional scan-lens/galvo-mirror combination, the two-mirror scanning geometry offers higher light efficiency and a more compact footprint, which could be beneficial to all OPM designs regardless of the use of a commercial base or not.
在这项工作中,我们提出了一种斜平面显微镜,其设计可与市售显微镜底座无缝配合。为了支持显微镜底座提供的所有功能(其中物镜的位置并不固定),我们采用了双镜扫描几何结构,可以在显微镜常规操作过程中补偿物镜位置的变化。我们的研究表明,在 100 倍、1.35 NA 物镜偏离设计位置 ± 1 mm 的位移范围内,PSF 尺寸在横向和轴向分别增加了 3% 和 11%,而在焦平面周围 ± 10 µm 的范围内,放大率误差为 0.5%。与更传统的扫描镜头/巨像镜组合相比,双镜扫描几何具有更高的光效和更紧凑的占地面积,这对所有 OPM 设计都有好处,无论是否使用商用底座。
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引用次数: 0
Terahertz polarimetric imaging of biological tissue: Monte Carlo modeling of signal contrast mechanisms due to Mie scattering 生物组织的太赫兹偏振成像:米氏散射导致的信号对比机制的蒙特卡罗建模
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-13 DOI: 10.1364/boe.515623
Kuangyi Xu and M. Hassan Arbab
Many promising biomedical applications have been proposed for terahertz (THz) spectroscopy and diagnostic imaging techniques. Polarimetric imaging systems are generally useful for enhancing imaging contrasts, yet the interplay between THz polarization changes and the random discrete structures in biological samples is not well understood. In this work, we performed Monte Carlo simulations of the propagation of polarized THz waves in skin and adipose tissues based on the Mie scattering from intrinsic structures, such as hair follicles or sweat glands. We show that the polarimetric contrasts are distinctly affected by concentration, size and dielectric properties of the scatterers, as well as the frequency and polarization of the incident THz waves. We describe the experimental requirements for observing and extracting these polarimetric signals due to the low energy and small angular spread of the back-scattered THz radiation. We analyzed the spatially integrated Mueller matrices of samples in the normal-incidence back-scattering geometry. We show that the frequency-dependent degree of polarization (DOP) can be used to infer the concentrations and dielectric contents of the scattering structures. Our modeling approach can be used to inform the design of the imaging modalities and the interpretation of the spectroscopic data in future terahertz biomedical imaging applications.
太赫兹(THz)光谱学和诊断成像技术已提出了许多前景广阔的生物医学应用。偏振成像系统通常有助于增强成像对比度,但人们对太赫兹偏振变化与生物样本中随机离散结构之间的相互作用还不甚了解。在这项工作中,我们根据毛囊或汗腺等固有结构的米氏散射,对极化太赫兹波在皮肤和脂肪组织中的传播进行了蒙特卡罗模拟。我们的研究表明,散射体的浓度、大小和介电性质,以及入射太赫兹波的频率和极化,都会明显影响极化对比度。我们描述了观测和提取这些极坐标信号的实验要求,因为后向散射的太赫兹辐射能量低、角度展宽小。我们分析了正常入射背向散射几何中样品的空间积分穆勒矩阵。我们表明,随频率变化的极化度 (DOP) 可用来推断散射结构的浓度和介电含量。在未来的太赫兹生物医学成像应用中,我们的建模方法可用于成像模式的设计和光谱数据的解释。
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引用次数: 0
Fast volumetric multifocus structured illumination microscopy of subcellular dynamics in living cells 活细胞亚细胞动态的快速容积多焦结构照明显微技术
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-11 DOI: 10.1364/boe.516261
Maximilian Lukas Senftleben, Antone Bajor, Eduardo Hirata, Sara Abrahamsson, and Hjalmar Brismar
Studying the nanoscale dynamics of subcellular structures is possible with 2D structured illumination microscopy (SIM). The method allows for acquisition with improved resolution over typical widefield. For 3D samples, the acquisition speed is inherently limited by the need to acquire sequential two-dimensional planes to create a volume. Here, we present a development of multifocus SIM designed to provide high volumetric frame rate by using fast synchronized electro-optical components. We demonstrate the high volumetric imaging capacity of the microscope by recording the dynamics of microtubule and endoplasmatic reticulum in living cells at up to 2.3 super resolution volumes per second for a total volume of 30 × 30 × 1.8 µm3.
利用二维结构照明显微镜(SIM)可以研究亚细胞结构的纳米级动态。与典型的宽场相比,该方法可提高采集分辨率。对于三维样品,由于需要连续采集二维平面来创建体积,因此采集速度受到固有限制。在此,我们介绍了多焦点 SIM 的开发情况,其设计目的是通过使用快速同步的电子光学元件来提供高容积帧频。我们通过记录活细胞中微管和内质网的动态,以每秒高达 2.3 个超分辨率体积(总体积为 30 × 30 × 1.8 µm3)的速度,展示了该显微镜的高体积成像能力。
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引用次数: 0
Low-cost 3D printed lenses for brightfield and fluorescence microscopy 用于明视野和荧光显微镜的低成本 3D 打印透镜
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-07 DOI: 10.1364/boe.514653
Jay Christopher, Liam M. Rooney, Mark Donnachie, Deepak Uttamchandani, Gail McConnell, and Ralf Bauer
We present the fabrication and implementation of low-cost optical quality 3D printed lenses, and their application as microscope objectives with different prescriptions. The imaging performance of the 3D printed lenses was benchmarked against commercially available optics including a 20 mm focal length 12.7 mm diameter NBK-7 plano-convex lens used as a low magnification objective, and a separate high magnification objective featuring three 6 mm diameter NBK-7 lenses with different positive and negative focal lengths. We describe the design and manufacturing processes to produce high-quality 3D printed lenses. We tested their surface quality using a stylus profilometer, showing that they conform to that of commercial glass counterpart lenses. The 3D printed lenses were used as microscope objectives in both brightfield and epi-fluorescence imaging of specimens including onion, cyanobacteria, and variegated Hosta leaves, demonstrating a sub-cellular resolution performance obtained with low-cost 3D printed optical elements within brightfield and fluorescence microscopy.
我们介绍了低成本光学质量三维打印透镜的制造和实施,以及将其用作不同度数显微镜物镜的情况。我们将三维打印透镜的成像性能与市售光学器件进行了比较,其中包括一个用作低倍物镜的 20 毫米焦距、直径 12.7 毫米的 NBK-7 平凸透镜,以及一个单独的高倍物镜,该物镜具有三个直径 6 毫米、正负焦距不同的 NBK-7 透镜。我们介绍了生产高质量 3D 打印透镜的设计和制造工艺。我们使用测针轮廓仪测试了镜片的表面质量,结果表明它们与商用玻璃镜片一致。我们将三维打印透镜用作明视野和外延荧光成像的显微镜物镜,对洋葱、蓝藻和变色玉簪叶片等标本进行了成像,证明了低成本三维打印光学元件在明视野和荧光显微镜中获得的亚细胞分辨率性能。
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引用次数: 0
Fair non-contact blood pressure estimation using imaging photoplethysmography 利用成像光压计进行公平的非接触式血压估算
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-05 DOI: 10.1364/boe.514241
Hongli Fang, Jiping Xiong, and Linying He
Hypertension is typically manifested as a latent symptom that requires detection through specialized equipment. This poses an inconvenience for individuals who need to undergo long-term blood pressure monitoring in their daily lives. Therefore, there is a need for a portable, non-contact method for estimating blood pressure. However, current non-contact blood pressure estimation methods often rely on relatively narrow datasets, lacking a broad range of blood pressure distributions. Additionally, their applicability is confined to controlled experimental environments. This study proposes a non-contact blood pressure estimation method suitable for various life scenarios, encompassing multiple age groups, diverse ethnicities, and individuals with different skin tones. The aim is to enhance the practicality and accuracy of existing non-contact blood pressure estimation methods. The research extracts the imaging photoplethysmogram (IPPG) signal from facial videos and processes the signal through four layers of filtering operations to obtain an IPPG signal reflecting pulse wave variations. A CNN+BiLSTM+GRU network structure is constructed to improve the accuracy of current non-contact blood pressure estimation methods. In comparison to existing approaches, the mean absolute error (MAE) for systolic blood pressure (SBP) and diastolic blood pressure (DBP) is reduced by 13.6% and 16.4%, respectively.
高血压通常表现为潜伏症状,需要通过专业设备进行检测。这给需要在日常生活中长期接受血压监测的人带来了不便。因此,需要一种便携式、非接触式的血压估测方法。然而,目前的非接触式血压估算方法往往依赖于相对狭窄的数据集,缺乏广泛的血压分布范围。此外,它们的适用性仅限于受控实验环境。本研究提出了一种适用于各种生活场景的非接触式血压估算方法,涵盖多个年龄组、不同种族和不同肤色的个体。目的是提高现有非接触式血压估算方法的实用性和准确性。该研究从面部视频中提取成像血压图(IPGP)信号,并通过四层滤波操作对信号进行处理,以获得反映脉搏波变化的 IPPGP 信号。通过构建 CNN+BiLSTM+GRU 网络结构,提高了现有非接触式血压估算方法的准确性。与现有方法相比,收缩压(SBP)和舒张压(DBP)的平均绝对误差(MAE)分别降低了 13.6% 和 16.4%。
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引用次数: 0
Performance enhancement of diffuse fluorescence tomography based on an extended Kalman filtering-long short term memory neural network correction model 基于扩展卡尔曼滤波-长短期记忆神经网络校正模型的漫反射荧光断层成像性能增强技术
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-04 DOI: 10.1364/boe.514041
Lingxiu Xing, Limin Zhang, Wenjing Sun, Zhuanxia He, Yanqi Zhang, and Feng Gao
To alleviate the ill-posedness of diffuse fluorescence tomography (DFT) reconstruction and improve imaging quality and speed, a model-derived deep-learning method is proposed by combining extended Kalman filtering (EKF) with a long short term memory (LSTM) neural network, where the iterative process parameters acquired by implementing semi-iteration EKF (SEKF) served as inputs to the LSTM neural network correction model for predicting the optimal fluorescence distributions. To verify the effectiveness of the SEKF-LSTM algorithm, a series of numerical simulations, phantom and in vivo experiments are conducted, and the experimental results are quantitatively evaluated and compared with the traditional EKF algorithm. The simulation experimental results show that the proposed new algorithm can effectively improve the reconstructed image quality and reconstruction speed. Importantly, the LSTM correction model trained by the simulation data also obtains satisfactory results in the experimental data, suggesting that the SEKF-LSTM algorithm possesses strong generalization ability and great potential for practical applications.
为了缓解弥散荧光断层成像(DFT)重建的假定性,提高成像质量和速度,本文提出了一种模型衍生的深度学习方法,将扩展卡尔曼滤波(EKF)与长短期记忆(LSTM)神经网络相结合,将半迭代EKF(SEKF)获得的迭代过程参数作为LSTM神经网络校正模型的输入,用于预测最佳荧光分布。为了验证 SEKF-LSTM 算法的有效性,研究人员进行了一系列数值模拟、幻影和活体实验,并对实验结果进行了定量评估,并与传统的 EKF 算法进行了比较。模拟实验结果表明,所提出的新算法能有效提高重建图像的质量和重建速度。重要的是,由仿真数据训练的 LSTM 修正模型在实验数据中也获得了令人满意的结果,表明 SEKF-LSTM 算法具有很强的泛化能力和巨大的实际应用潜力。
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引用次数: 0
Analog multiplexing of a laser clock and computational photon counting for fast fluorescence lifetime imaging microscopy 用于快速荧光寿命成像显微镜的激光时钟模拟多路复用和计算光子计数技术
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-04 DOI: 10.1364/boe.514813
Rishyashring R. Iyer, Janet E. Sorrells, Kevin K. D. Tan, Lingxiao Yang, Geng Wang, Haohua Tu, and Stephen A. Boppart
The dynamic range and fluctuations of fluorescence intensities and lifetimes in biological samples are large, demanding fast, precise, and versatile techniques. Among the high-speed fluorescence lifetime imaging microscopy (FLIM) techniques, directly sampling the output of analog single-photon detectors at GHz rates combined with computational photon counting can handle a larger range of photon rates. Traditionally, the laser clock is not sampled explicitly in fast FLIM; rather the detection is synchronized to the laser clock so that the excitation pulse train can be inferred from the cumulative photon statistics of several pixels. This has two disadvantages for sparse or weakly fluorescent samples: inconsistencies in inferring the laser clock within a frame and inaccuracies in aligning the decay curves from different frames for averaging. The data throughput is also very inefficient in systems with repetition rates much larger than the fluorescence lifetime due to significant silent regions where no photons are expected. We present a method for registering the photon arrival times to the excitation using time-domain multiplexing for fast FLIM. The laser clock is multiplexed with photocurrents into the silent region. Our technique does not add to the existing data bottleneck, has the sub-nanosecond dead time of computational photon counting based fast FLIM, works with various detectors, lasers, and electronics, and eliminates the errors in lifetime estimation in photon-starved conditions. We demonstrate this concept on two multiphoton setups of different laser repetition rates for single and multichannel FLIM multiplexed into a single digitizer channel for real-time imaging of biological samples.
生物样品中荧光强度和寿命的动态范围和波动很大,需要快速、精确和多功能的技术。在高速荧光寿命成像显微镜(FLIM)技术中,以 GHz 速率直接对模拟单光子探测器的输出进行采样并结合计算光子计数可以处理更大范围的光子速率。传统上,快速 FLIM 并不对激光时钟进行明确采样,而是将检测与激光时钟同步,这样就能从多个像素的累积光子统计中推断出激发脉冲序列。这对于稀疏或弱荧光样本来说有两个缺点:一是在一帧内推断激光时钟不一致,二是对不同帧的衰减曲线进行平均时不准确。在重复率远大于荧光寿命的系统中,数据吞吐量的效率也非常低,这是因为预计没有光子的静默区非常大。我们提出了一种利用时域多路复用技术将光子到达时间记录到激发上的方法,用于快速荧光成像。激光时钟与进入静默区的光电流复用。我们的技术不会增加现有的数据瓶颈,具有基于计算光子计数的快速 FLIM 的亚纳秒级死区时间,可与各种探测器、激光器和电子设备配合使用,并消除了光子饥饿条件下的寿命估计误差。我们在两个不同激光重复率的多光子装置上演示了这一概念,这两个装置分别用于单通道和多通道 FLIM,并复用到单个数字转换器通道,用于生物样本的实时成像。
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引用次数: 0
Optical coherence tomography for multicellular tumor spheroid category recognition and drug screening classification via multi-spatial-superficial-parameter and machine learning 通过多空间-表面参数和机器学习,利用光学相干断层扫描技术进行多细胞肿瘤球体类别识别和药物筛选分类
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-03-04 DOI: 10.1364/boe.514079
Feng Yan, Bornface Mutembei, Trisha Valerio, Gokhan Gunay, Ji-Hee Ha, Qinghao Zhang, Chen Wang, Ebenezer Raj Selvaraj Mercyshalinie, Zaid A. Alhajeri, Fan Zhang, Lauren E. Dockery, Xinwei Li, Ronghao Liu, Danny N. Dhanasekaran, Handan Acar, Wei R. Chen, and Qinggong Tang
Optical coherence tomography (OCT) is an ideal imaging technique for noninvasive and longitudinal monitoring of multicellular tumor spheroids (MCTS). However, the internal structure features within MCTS from OCT images are still not fully utilized. In this study, we developed cross-statistical, cross-screening, and composite-hyperparameter feature processing methods in conjunction with 12 machine learning models to assess changes within the MCTS internal structure. Our results indicated that the effective features combined with supervised learning models successfully classify OVCAR-8 MCTS culturing with 5,000 and 50,000 cell numbers, MCTS with pancreatic tumor cells (Panc02-H7) culturing with the ratio of 0%, 33%, 50%, and 67% of fibroblasts, and OVCAR-4 MCTS treated by 2-methoxyestradiol, AZD1208, and R-ketorolac with concentrations of 1, 10, and 25 µM. This approach holds promise for obtaining multi-dimensional physiological and functional evaluations for using OCT and MCTS in anticancer studies.
光学相干断层扫描(OCT)是对多细胞肿瘤球(MCTS)进行无创和纵向监测的理想成像技术。然而,从 OCT 图像中获得的多细胞肿瘤球内部结构特征仍未得到充分利用。在本研究中,我们开发了交叉统计、交叉筛选和复合超参数特征处理方法,并结合 12 个机器学习模型来评估多细胞肿瘤球内部结构的变化。结果表明,有效特征与监督学习模型相结合,成功地对培养细胞数为 5,000 和 50,000 的 OVCAR-8 MCTS、培养成纤维细胞比例为 0%、33%、50% 和 67% 的胰腺肿瘤细胞(Panc02-H7)MCTS 以及经 2-甲氧基雌二醇、AZD1208 和 R-Ketorolac 处理(浓度为 1、10 和 25 µM)的 OVCAR-4 MCTS 进行了分类。这种方法有望在抗癌研究中利用 OCT 和 MCTS 获得多维生理和功能评估。
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引用次数: 0
Exploring in-vivo infrared spectroscopy for nail-based diabetes screening 探索利用体内红外光谱进行指甲糖尿病筛查
IF 3.4 2区 医学 Q1 Physics and Astronomy Pub Date : 2024-02-28 DOI: 10.1364/boe.520102
Daniela Lazaro-Pacheco, Philip F Taday, and Päivi Maria Paldánius
Diabetes screening is traditionally complex, inefficient, and reliant on invasive sampling. This study evaluates near-infrared spectroscopy for non-invasive detection of glycated keratin in nails in vivo. Glycation of keratin, prevalent in tissues like nails and skin, is a key indicator of T2DM risk. In this study involving 200 participants (100 with diabetes, 100 without), NIR’s efficacy was compared against a point-of-care HbA1c analyzer. Results showed a specificity of 92.9% in diabetes risk assessment. This study highlights the proposed NIR system potential as a simple, reliable tool for early diabetes screening and risk management in various healthcare settings.
糖尿病筛查历来复杂、低效,而且依赖于侵入性采样。本研究评估了近红外光谱技术对体内指甲中糖化角蛋白的无创检测。角蛋白糖化普遍存在于指甲和皮肤等组织中,是 T2DM 风险的一个关键指标。在这项涉及 200 名参与者(100 名糖尿病患者,100 名非糖尿病患者)的研究中,近红外的功效与护理点 HbA1c 分析仪进行了比较。结果显示,近红外在糖尿病风险评估中的特异性高达 92.9%。这项研究凸显了近红外系统作为一种简单、可靠的工具,在各种医疗环境下进行早期糖尿病筛查和风险管理的潜力。
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引用次数: 0
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Biomedical optics express
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