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Stimulated Brillouin scattering flow cytometry. 受激布里渊散射流式细胞仪。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-25 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.537602
Jake R Rosvold, Giulia Zanini, Chenchen Handler, Eric Frank, Jiarui Li, Michele I Vitolo, Stuart S Martin, Giuliano Scarcelli

We present the use of stimulated Brillouin scattering spectroscopy to achieve rapid measurements of cell biomechanics in a flow cytometer setup. Specifically, our stimulated Brillouin scattering flow cytometry can acquire at a rate of 200 Hz, with a spectral acquisition time of 5 ms, which marks a 10x improvement compared to previous demonstrations of spontaneous Brillouin scattering flow cytometry. We experimentally validate our stimulated Brillouin scattering flow cytometer by measuring cell populations of normal breast epithelial cells and metastatic breast epithelial cancer cells.

我们介绍了在流式细胞仪设置中使用受激布里渊散射光谱快速测量细胞生物力学的方法。具体来说,我们的受激布里渊散射流式细胞仪能以 200 Hz 的速率采集,光谱采集时间为 5 毫秒,与之前展示的自发布里渊散射流式细胞仪相比,提高了 10 倍。我们通过测量正常乳腺上皮细胞和转移性乳腺上皮癌细胞的细胞群,对刺激布里渊散射流式细胞仪进行了实验验证。
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引用次数: 0
OCTA-ReVA: an open-source toolbox for comprehensive retinal vessel feature analysis in optical coherence tomography angiography. OCTA-ReVA:用于光学相干断层血管造影中视网膜血管特征综合分析的开源工具箱。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-25 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.537727
Albert K Dadzie, David Le, Mansour Abtahi, Behrouz Ebrahimi, Tobiloba Adejumo, Taeyoon Son, Michael J Heiferman, Jennifer I Lim, Xincheng Yao

Optical coherence tomography angiography (OCTA) has significantly advanced the study and diagnosis of eye diseases. However, current clinical OCTA systems and software tools lack comprehensive quantitative analysis capabilities, limiting their full clinical utility. This paper introduces the OCTA Retinal Vessel Analyzer (OCTA-ReVA), a versatile open-source platform featuring a user-friendly graphical interface designed for the automated extraction and quantitative analysis of OCTA features. OCTA-ReVA includes traditional established OCTA features based on binary vascular image processing, such as blood vessel density (BVD), foveal avascular zone area (FAZ-A), blood vessel tortuosity (BVT), and blood vessel caliber (BVC). Additionally, it introduces new features based on blood perfusion intensity processing, such as perfusion intensity density (PID), vessel area flux (VAF), and normalized blood flow index (NBFI), which provide deeper insights into retinal perfusion conditions. These additional capabilities are crucial for the early detection and monitoring of retinal diseases. OCTA-ReVA demystifies the intricate task of retinal vasculature quantification, offering a robust tool for researchers and clinicians to objectively evaluate eye diseases and enhance the precision of retinal health assessments.

光学相干断层血管成像(OCTA)极大地推动了眼科疾病的研究和诊断。然而,目前的临床 OCTA 系统和软件工具缺乏全面的定量分析能力,限制了其临床应用的充分发挥。本文介绍了 OCTA 视网膜血管分析仪(OCTA-ReVA),这是一个多功能开源平台,具有用户友好的图形界面,专为自动提取和定量分析 OCTA 特征而设计。OCTA-ReVA 包括基于二元血管图像处理的传统 OCTA 特征,如血管密度 (BVD)、眼窝无血管区面积 (FAZ-A)、血管迂曲度 (BVT) 和血管口径 (BVC)。此外,它还引入了基于血液灌注强度处理的新功能,如灌注强度密度 (PID)、血管面积通量 (VAF) 和归一化血流指数 (NBFI),从而更深入地了解视网膜灌注状况。这些附加功能对于早期检测和监测视网膜疾病至关重要。OCTA-ReVA 揭开了视网膜血管量化这一复杂任务的神秘面纱,为研究人员和临床医生客观评估眼部疾病和提高视网膜健康评估的精确度提供了强有力的工具。
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引用次数: 0
Single-shot quantitative phase microscopy: a multi-functional tool for cell analysis. 单次定量相显微镜:细胞分析的多功能工具。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-24 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.533091
Ana Espinosa-Momox, Brandon Norton, Maria Cywinska, Bryce Evans, Juan Vivero-Escoto, Rosario Porras-Aguilar

This study showcases the multifunctionality of a single-shot quantitative phase microscopy (QPM) system for comprehensive cell analysis. The system captures four high-contrast images in one shot, enabling tasks like cell segmentation, measuring cell confluence, and estimating cell mass. We demonstrate the usability of the QPM system in routine biological workflows, showing how its integration with computational algorithms enables automated, precise analysis, achieving accuracy scores between 85% and 97% across samples with varying cell densities, even those with low signal-to-noise ratios. This cost-effective tool operates under low-intensity light and resists vibrations, making it highly versatile for researchers in both optical and biological fields.

这项研究展示了用于细胞综合分析的单次定量相显微镜(QPM)系统的多功能性。该系统一次拍摄四幅高对比度图像,可完成细胞分割、测量细胞汇合度和估计细胞质量等任务。我们演示了 QPM 系统在常规生物工作流程中的可用性,展示了该系统与计算算法的集成如何实现自动、精确的分析,在不同细胞密度的样本中,甚至在信噪比较低的样本中,准确率达到 85% 到 97%。这种高性价比的工具可在低强度光线下工作,并能抵抗振动,因此对光学和生物领域的研究人员来说都非常灵活。
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引用次数: 0
Development of a carboxymethyl chitosan functionalized slide for small molecule detection using oblique-incidence reflectivity difference technology. 利用斜入射反射率差技术开发用于小分子检测的羧甲基壳聚糖功能化载玻片。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-19 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.534563
Mengjing Xu, Boyang Shi, Haofeng Li, Xiaohan Mai, Lan Mi, Jiong Ma, Xiangdong Zhu, Guowei Wang, Yiyan Fei

Label-free optical biosensors have become powerful tools in the study of biomolecular interactions without the need for labels. High throughput and low detection limit are desirable for rapid and accurate biomolecule detection. The oblique-incidence reflectivity difference (OI-RD) technique is capable of detecting thousands of biomolecular interactions in a high-throughput mode, specifically for biomolecules larger than 1000 Da. In order to enhance the detection capability of OI-RD for small molecules (typically < 500 Da), we have developed a three-dimensional biochip that utilized carboxymethyl chitosan (CMCS) functionalized slides. By investigating various factors such as sonication time, protein immobilization time, CMCS molecular weight, and glutaraldehyde (GA) functionalization time, we have achieved a detection limit of 6.8 pM for avidin (68 kDa). Furthermore, accurate detection of D-biotin with a molecular weight of 244 Da has also been achieved. This paper presents an effective solution for achieving both high throughput and low detection limits using the OI-RD technique in the field of biomolecular interaction detection.

无标记光学生物传感器已成为研究生物分子相互作用的强大工具,无需标记。高通量和低检测限是快速准确检测生物分子的理想选择。斜入射反射率差分(OI-RD)技术能够在高通量模式下检测成千上万的生物分子相互作用,尤其适用于大于 1000 Da 的生物分子。为了提高 OI-RD 对小分子(通常为
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引用次数: 0
DMD and microlens array as a switchable module for illumination angle scanning in optical diffraction tomography. 将 DMD 和微透镜阵列作为光学衍射断层扫描中照明角扫描的可切换模块。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-18 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.535123
Siqi Yang, Jeongsoo Kim, Mary E Swartz, Johann K Eberhart, Shwetadwip Chowdhury

Optical diffraction tomography (ODT) enables label-free and morphological 3D imaging of biological samples using refractive-index (RI) contrast. To accomplish this, ODT systems typically capture multiple angular-specific scattering measurements, which are used to computationally reconstruct a sample's 3D RI. Standard ODT systems employ scanning mirrors to generate angular illuminations. However, scanning mirrors are limited to illuminating the sample from only one angle at a time. Furthermore, when operated at high speeds, these mirrors may exhibit mechanical instabilities that compromise image quality and measurement speed. Recently, newer ODT systems have been introduced that utilize digital-micromirror devices (DMD), spatial light modulators (SLMs), or LED arrays to achieve switchable angle-scanning with no physically-scanning components. However, these systems associate with power inefficiencies and/or spurious diffraction orders that can also limit imaging performance. In this work, we developed a novel non-interferometric ODT system that utilizes a fully switchable module for angle scanning composed of a DMD and microlens array (MLA). Compared to other switchable ODT systems, this module enables each illumination angle to be generated fully independently from every other illumination angle (i.e., no spurious diffraction orders) while also optimizing the power efficiency based on the required density of illumination angles. We validate the quantitative imaging capability of this system using calibration microspheres. We also demonstrate its capability for imaging multiple-scattering samples by imaging an early-stage zebrafish embryo.

光学衍射层析(ODT)可利用折射率(RI)对比对生物样本进行无标记和形态学三维成像。为了实现这一目标,ODT 系统通常会捕捉多个特定角度的散射测量值,用于计算重建样品的三维 RI。标准 ODT 系统采用扫描镜产生角度照明。然而,扫描镜每次只能从一个角度照射样品。此外,在高速运行时,这些反射镜可能会出现机械不稳定性,从而影响图像质量和测量速度。最近,新推出的 ODT 系统利用数字微镜设备 (DMD)、空间光调制器 (SLM) 或 LED 阵列来实现可切换角度扫描,而无需物理扫描组件。然而,这些系统存在功率效率低下和/或虚假衍射阶次的问题,也会限制成像性能。在这项工作中,我们开发了一种新型非干涉式 ODT 系统,该系统利用由 DMD 和微透镜阵列 (MLA) 组成的完全可切换模块进行角度扫描。与其他可切换式 ODT 系统相比,该模块可使每个照明角度的产生完全独立于其他照明角度(即无杂散衍射阶),同时还可根据所需的照明角度密度优化功率效率。我们利用校准微球验证了该系统的定量成像能力。我们还通过对早期斑马鱼胚胎成像,证明了该系统对多重散射样本成像的能力。
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引用次数: 0
Investigating the effect of limited spectral information on NIRS-derived changes in hemoglobin and cytochrome-c-oxidase concentration with a diffusion-based model. 利用基于扩散的模型研究有限光谱信息对近红外光谱衍生的血红蛋白和细胞色素-c-氧化酶浓度变化的影响。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-17 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.531775
Georgina Leadley, Robert J Cooper, Topun Austin, Jeremy C Hebden, Gemma Bale

This paper investigates the theoretical capability of near-infrared spectroscopy (NIRS) systems to accurately measure changes in the oxidation state of cerebral cytochrome-c-oxidase (CCO) alongside the hemoglobins, for a deeper understanding of NIRS limitations. Concentration changes of oxy and deoxyhemoglobin (HbO and HbR) indicate the oxygen status of blood vessels and correlate with several other physiological parameters across different pathologies. The oxidation state of CCO indicates cellular energy usage efficiency through oxidative metabolism, potentially serving as a biomarker for brain and other tissue disorders. This study employs an analytical model based on the diffusion equation and statistical analyses to explore the dependency of estimated concentration changes on various systematic parameters, such as choice of wavelengths, spectral bandwidth, and uncertainties in extinction coefficient (ε) and differential pathlength factor (DPF). When there is a 10% uncertainty in DPF and ε, errors were found to be highly dependent on the number of discrete wavelengths, but not on their bandwidth if appropriate considerations are taken to account for it.

本文研究了近红外光谱(NIRS)系统精确测量脑细胞色素-c-氧化酶(CCO)氧化状态变化以及血红蛋白的理论能力,以便更深入地了解 NIRS 的局限性。氧血红蛋白和脱氧血红蛋白(HbO 和 HbR)的浓度变化表明血管的氧状态,并与不同病理情况下的其他几个生理参数相关。CCO 的氧化状态表明细胞通过氧化代谢利用能量的效率,有可能成为大脑和其他组织疾病的生物标志物。本研究采用基于扩散方程的分析模型和统计分析来探讨估计浓度变化对各种系统参数的依赖性,如波长选择、光谱带宽以及消光系数(ε)和微分路径长度因子(DPF)的不确定性。当 DPF 和 ε 的不确定性为 10%时,误差高度依赖于离散波长的数量,但如果考虑到适当的带宽,误差则与带宽无关。
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引用次数: 0
Age dependence of the average refractive index of the isolated human crystalline lens. 离体人类晶状体平均折射率的年龄依赖性。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-17 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.536501
Ramya Natarajan, Bianca Maceo Heilman, Marco Ruggeri, Arthur Ho, Vivek M Singh, Robert Augusteyn, Jean-Marie Parel, Pravin K Vaddavalli, Fabrice Manns

We measured the average group refractive index (RI) of 120 isolated lenses from 120 human donors (age: 0.03 to 61 years). The average group RI was calculated from a measurement of the optical thickness of the lens using optical coherence tomography and the apparent window shift of the test chamber caused by the lens. The estimated measurement uncertainty was ±0.004. The group RI at 880 nm was converted to phase RI at 589 nm using the dispersion equation of water and protein. From 2 to 61 years, the mean value of the RI was 1.415 ± 0.002 (group index at 880 nm) and 1.406 ± 0.002 (phase index at 589 nm) independent of age (p = 0.774). Two lenses from donors of age 0.33 and 3 months had significantly lower RI (group index: 1.405 and 1.403; phase index: 1.396 and 1.394). From age 2 to 61, the average lens RI is constant with age within the measurement uncertainty (±0.004).

我们测量了来自 120 名人体捐献者(年龄:0.03 至 61 岁)的 120 个离体晶状体的平均组折射率 (RI)。通过光学相干断层扫描测量镜片的光学厚度以及镜片引起的测试室视窗偏移,计算出平均组别 RI。估计测量不确定性为 ±0.004。利用水和蛋白质的色散方程,将 880 纳米波长的组 RI 转换为 589 纳米波长的相 RI。从 2 岁到 61 岁,RI 的平均值为 1.415 ± 0.002(880 纳米波长的组指数)和 1.406 ± 0.002(589 纳米波长的相指数),与年龄无关(p = 0.774)。年龄分别为 0.33 个月和 3 个月的两个供体的镜片的 RI 明显较低(组指数:1.405 和 1.403;相指数:1.396 和 1.394)。从 2 岁到 61 岁,镜片平均 RI 随年龄的变化而变化,测量误差在 ±0.004 范围内。
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引用次数: 0
Fluorescence super-resolution microscopy via fluctuation-based multi-route synergy. 通过基于波动的多路协同技术实现荧光超分辨率显微镜。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-16 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.534067
Zhiping Zeng, Biqing Xu, Jin Qiu, Xinyi Chen, Yantang Huang, Canhua Xu

Fluorescence fluctuation-based super-resolution microscopy (FF-SRM) is an economical and widely applicable technique that significantly enhances the spatial resolution of fluorescence imaging by capitalizing on fluorescence intermittency. However, each variant of FF-SRM imaging has inherent limitations. This study proposes a super-resolution reconstruction strategy (synSRM) by synergizing multiple variants of the FF-SRM approach to address the limitations and achieve high-quality and high-resolution imaging. The simulation and experimental results demonstrate that, compared to images reconstructed using single FF-SRM algorithms, by selecting suitable synSRM routes according to various imaging conditions, further improvements of the spatial resolution and image reconstruction quality can be obtained for super-resolution fluorescence imaging.

基于荧光波动的超分辨显微镜(FF-SRM)是一种经济而又广泛应用的技术,它利用荧光的间歇性显著提高了荧光成像的空间分辨率。然而,FF-SRM 成像的每种变体都有其固有的局限性。本研究提出了一种超分辨率重建策略(synSRM),通过协同 FF-SRM 方法的多个变体来解决局限性问题,实现高质量、高分辨率成像。模拟和实验结果表明,与使用单一 FF-SRM 算法重建的图像相比,根据不同的成像条件选择合适的 synSRM 路线,可以进一步提高超分辨率荧光成像的空间分辨率和图像重建质量。
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引用次数: 0
TDT-MIL: a framework with a dual-channel spatial positional encoder for weakly-supervised whole slide image classification. TDT-MIL:双通道空间位置编码器弱监督整张幻灯片图像分类框架。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-13 eCollection Date: 2024-10-01 DOI: 10.1364/BOE.530534
Hongbin Zhang, Ya Feng, Jin Zhang, Guangli Li, Jianguo Wu, Donghong Ji

The classic multiple instance learning (MIL) paradigm is harnessed for weakly-supervised whole slide image (WSI) classification. The spatial position relationship located between positive tissues is crucial for this task due to the small percentage of these tissues in billions of pixels, which has been overlooked by most studies. Therefore, we propose a framework called TDT-MIL. We first serially connect a convolutional neural network and transformer for basic feature extraction. Then, a novel dual-channel spatial positional encoder (DCSPE) module is designed to simultaneously capture the complementary local and global positional information between instances. To further supplement the spatial position relationship, we construct a convolutional triple-attention (CTA) module to attend to the inter-channel information. Thus, the spatial positional and inter-channel information is fully mined by our model to characterize the key pathological semantics in WSI. We evaluated TDT-MIL on two publicly available datasets, including CAMELYON16 and TCGA-NSCLC, with the corresponding classification accuracy and AUC up to 91.54%, 94.96%, and 90.21%, 94.36%, respectively, outperforming state-of-the-art baselines. More importantly, our model possesses a satisfactory capability in solving the imbalanced WSI classification task using an ingenious but interpretable structure.

经典的多实例学习(MIL)范式被用于弱监督全切片图像(WSI)分类。由于阳性组织在数十亿像素中所占比例较小,因此位于阳性组织之间的空间位置关系对这项任务至关重要,而大多数研究都忽略了这一点。因此,我们提出了一个名为 TDT-MIL 的框架。我们首先将卷积神经网络和变压器串联起来,进行基本的特征提取。然后,我们设计了一个新颖的双通道空间位置编码器(DCSPE)模块,以同时捕捉实例之间互补的局部和全局位置信息。为了进一步补充空间位置关系,我们构建了一个卷积三重关注(CTA)模块来关注通道间信息。因此,我们的模型可以充分挖掘空间位置和信道间信息,从而描述 WSI 中的关键病理语义。我们在两个公开数据集(包括 CAMELYON16 和 TCGA-NSCLC)上对 TDT-MIL 进行了评估,其相应的分类准确率和 AUC 分别高达 91.54%、94.96% 和 90.21%、94.36%,优于最先进的基线模型。更重要的是,我们的模型具有令人满意的能力,能利用巧妙而可解释的结构解决不平衡的 WSI 分类任务。
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引用次数: 0
Chirp excitation for natural frequency optical coherence elastography 用于固有频率光学相干弹性成像的啁啾激励
IF 3.4 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-09-09 DOI: 10.1364/boe.536685
Chengjin Song, Weichao He, Pengfei Song, Jinping Feng, Yanping Huang, Jingjiang Xu, Lin An, Jia Qin, Kai Gao, Michael D. Twa, Gongpu Lan
Optical coherence elastography (OCE) has recently been used to characterize the natural frequencies of delicate tissues (e.g., the in vivo human cornea) with sub-micron tissue oscillation magnitudes. Here, we investigate broadband spectrum sample stimulation using a contact-based piezoelectric transducer (PZT) chirp excitation and compare its performance with a non-contact, air-pulse excitation for OCE measurements on 1.0-7.5% agar phantoms and an ex vivo porcine cornea under intraocular pressures (IOPs) of 5-40 mmHg. The 3-ms duration air-pulse generated a ∼0–840 Hz excitation spectrum, effectively quantifying the first-order natural frequencies in softer samples (e.g., 1.0%–4.0% agar: 239–782 Hz, 198 Hz/%; porcine cornea: 68–414 Hz, 18 Hz/mmHg, IOP: 5–25 mmHg), but displayed limitations in measuring natural frequencies for stiffer samples (e.g., 4.5%–7.5% agar, porcine cornea: IOP ≥ 30 mmHg) or higher order natural frequency components. In contrast, the chirp excitation produced a much wider spectrum (e.g., 0–5000 Hz), enabling the quantification of both first-order natural frequencies (1.0%–7.5% agar: 253–1429 Hz, 181 Hz/%; porcine cornea: 76–1240 Hz, 32 Hz/mmHg, IOP: 5–40 mmHg) and higher order natural frequencies. A modified Bland-Altman analysis (mean versus relative difference in natural frequency) showed a bias of 20.4%, attributed to the additional mass and frequency introduced by the contact nature of the PZT probe. These findings, especially the advantages and limitations of both excitation methods, can be utilized to validate the potential application of natural frequency OCE, paving the way for the ongoing development of biomechanical characterization methods utilizing sub-micron tissue oscillation features.
光学相干弹性成像(OCE)最近已被用于表征脆弱组织(如活体人类角膜)的自然频率,其组织振荡幅度可达亚微米级。在此,我们研究了使用接触式压电换能器(PZT)啁啾激励的宽带频谱样本刺激,并比较了其与非接触式空气脉冲激励在眼内压(IOP)为 5-40 mmHg 的情况下对 1.0-7.5% 琼脂模型和活体猪角膜进行 OCE 测量的性能。持续时间为 3 毫秒的空气脉冲产生了 ∼0-840 Hz 的激励频谱,有效地量化了较软样品(例如,1.0%-4.0%)的一阶固有频率、1.0%-4.0%琼脂:239-782 Hz,198 Hz/%;猪角膜:68-414 Hz,18 Hz/mmHg,眼压:5-25 mmHg),但在测量较硬样品(如 4.5%-7.5%琼脂,猪角膜:眼压≥ 30 mmHg)的自然频率或高阶自然频率成分时显示出局限性。相比之下,啁啾激励产生的频谱要宽得多(如 0-5000 Hz),能够量化一阶固有频率(1.0%-7.5% 琼脂:253-1429 Hz,181 Hz/%;猪角膜:76-1240 Hz,32 Hz/mmHg,眼压:5-40 mmHg)和高阶固有频率。修正的布兰-阿尔特曼分析(自然频率的平均值与相对差异)显示偏差为 20.4%,这是由于 PZT 探头的接触性质带来了额外的质量和频率。这些发现,特别是两种激励方法的优势和局限性,可以用来验证自然频率 OCE 的潜在应用,为正在进行的利用亚微米组织振荡特征的生物力学表征方法的开发铺平道路。
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引用次数: 0
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Biomedical optics express
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