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Pixel-level classification of pigmented skin cancer lesions using multispectral autofluorescence lifetime dermoscopy imaging 利用多光谱自发荧光终生皮肤镜成像对色素性皮肤癌病变进行像素级分类
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-02 DOI: 10.1364/boe.523831
Priyanka Vasanthakumari, Renan Romano, Ramon Rosa, A. Salvio, Vladislav V. Yakovlev, Cristina Kurachi, Jason Hirshburg, Javier A Jo
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引用次数: 0
Non-invasive laser speckle contrast imaging (LSCI) of extra-embryonic blood vessels in intact avian eggs at early developmental stages 早期发育阶段完整禽蛋胚外血管的无创激光斑点对比成像(LSCI)
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-01 DOI: 10.1364/boe.530366
Zhenyu Dong, Simon Mahler, Carol Readhead, Xi Chen, Maya Dickson, Marianne Bronner, Changhuei Yang
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引用次数: 0
Adaptive optics in an oblique plane microscope 斜面显微镜中的自适应光学技术
IF 3.4 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-28 DOI: 10.1364/boe.524013
Conor Mcfadden, Zach Marin, Bingying Chen, Stephan Daetwyler, Xiaoding Wang, Divya Rajendran, Kevin M. Dean, Reto Fiolka
Adaptive optics (AO) can restore diffraction-limited performance when imaging beyond superficial cell layers in vivo and in vitro, and as such, is of interest for advanced 3D microscopy methods such as light-sheet fluorescence microscopy (LSFM). In a typical LSFM system, the illumination and detection paths are separate and subject to different optical aberrations. To achieve optimal microscope performance, it is necessary to sense and correct these aberrations in both light paths, resulting in a complex microscope system. Here, we show that in an oblique plane microscope (OPM), a type of LSFM with a single primary objective lens, the same deformable mirror can correct both illumination and fluorescence detection. Besides reducing the complexity, we show that AO in OPM also restores the relative alignment of the light-sheet and focal plane, and that a projection imaging mode can stabilize and improve the wavefront correction in a sensorless AO format. We demonstrate OPM with AO on fluorescent nanospheres and by imaging the vasculature and cancer cells in zebrafish embryos embedded in a glass capillary, restoring diffraction limited resolution and improving the signal strength twofold.
自适应光学(AO)可以恢复体内和体外浅表层细胞成像时的衍射限制性能,因此对光片荧光显微镜(LSFM)等先进的三维显微方法很有意义。在典型的 LSFM 系统中,照明和检测路径是分开的,并受到不同光学像差的影响。为了获得最佳的显微镜性能,有必要感测和校正两条光路中的这些像差,从而形成一个复杂的显微镜系统。在这里,我们展示了在斜面显微镜(OPM)(一种具有单主物镜的 LSFM)中,同一可变形镜可以同时校正照明和荧光检测。除了降低复杂性外,我们还展示了 OPM 中的自动光学还能恢复光片和焦平面的相对对齐,而且投影成像模式可以稳定和改进无传感器自动光学格式中的波前校正。我们演示了在荧光纳米球上使用 AO 的 OPM,并通过对嵌入玻璃毛细管中的斑马鱼胚胎的血管和癌细胞进行成像,恢复了衍射受限的分辨率,并将信号强度提高了两倍。
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引用次数: 0
3D super-resolution optical fluctuation imaging with temporal focusing two-photon excitation. 利用时间聚焦双光子激发的三维超分辨率光学波动成像。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-28 eCollection Date: 2024-07-01 DOI: 10.1364/BOE.523430
Pawel Szczypkowski, Monika Pawlowska, Radek Lapkiewicz

3D super-resolution fluorescence microscopy typically requires sophisticated setups, sample preparation, or long measurements. A notable exception, SOFI, only requires recording a sequence of frames and no hardware modifications whatsoever but being a wide-field method, it faces problems in thick, dense samples. We combine SOFI with temporal focusing two-photon excitation - the wide-field method that is capable of exciting a thin slice in 3D volume. Temporal focusing is simple to implement whenever the excitation path of the microscope can be accessed. The implementation of SOFI is straightforward. By merging these two methods, we obtain super-resolved 3D images of neurons stained with quantum dots. Our approach offers reduced bleaching of out-of-focus fluorescent probes and an improved signal-to-background ratio that can be used when robust resolution improvement is required in thick, dense samples.

三维超分辨率荧光显微镜通常需要复杂的设置、样品制备或长时间测量。但 SOFI 是一个显著的例外,它只需要记录一连串的帧,不需要对硬件进行任何修改,但作为一种宽视场方法,它在厚而密集的样品中面临着问题。我们将 SOFI 与时间聚焦双光子激发相结合,这种宽场方法能够激发三维体积中的薄片。只要能进入显微镜的激发路径,时间聚焦法就很容易实现。SOFI 的实施也很简单。通过合并这两种方法,我们获得了量子点染色神经元的超分辨三维图像。我们的方法减少了焦外荧光探针的漂白现象,提高了信噪比,可用于在厚而密集的样本中提高分辨率。
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引用次数: 0
In vivo imaging of human retinal ganglion cells using optical coherence tomography without adaptive optics 使用光学相干断层成像技术对人类视网膜神经节细胞进行活体成像,无需自适应光学技术
IF 3.4 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-28 DOI: 10.1364/boe.533249
Furu Zhang, Katherine Kovalick, Achyut Raghavendra, Somayyeh Soltanian-Zadeh, Sina Farsiu, Daniel X. Hammer, Zhuolin Liu
Retinal ganglion cells play an important role in human vision, and their degeneration results in glaucoma and other neurodegenerative diseases. Imaging these cells in the living human retina can greatly improve the diagnosis and treatment of glaucoma. However, owing to their translucent soma and tight packing arrangement within the ganglion cell layer (GCL), successful imaging has only been achieved with sophisticated research-grade adaptive optics (AO) systems. For the first time we demonstrate that GCL somas can be resolved and cell morphology can be quantified using non-AO optical coherence tomography (OCT) devices with optimal parameter configuration and post-processing.
视网膜神经节细胞在人类视觉中发挥着重要作用,它们的退化会导致青光眼和其他神经退行性疾病。对活体视网膜中的这些细胞进行成像可大大提高青光眼的诊断和治疗水平。然而,由于神经节细胞体是半透明的,而且在神经节细胞层(GCL)内排列紧密,因此只能通过精密的研究级自适应光学(AO)系统才能成功成像。我们首次证明,使用非自适应光学相干断层扫描(OCT)设备,通过最佳参数配置和后处理,可以分辨 GCL 体节并量化细胞形态。
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引用次数: 0
Coagulation depth estimation using a line scanner for depth-resolved laser speckle contrast imaging 利用线扫描仪进行深度分辨激光斑点对比成像的凝固深度估计
IF 3.4 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-27 DOI: 10.1364/boe.529043
Johannes D. Johansson, Martin Hultman, Rolf Saager
Partial-thickness burn wounds extend partially through the dermis, leaving many pain receptors intact and making the injuries very painful. Due to the painfulness, quick assessment of the burn depth is important to not delay surgery of the wound if needed. Laser speckle imaging (LSI) of skin blood flow can be helpful in finding severe coagulation zones with impaired blood flow. However, LSI measurements are typically too superficial to properly reach the full depth of the adult dermis and cannot resolve the flow in depth. Diffuse correlation spectroscopy (DCS) uses varying source-detector separations to allow differentiation of flow depths but requires time-consuming 2D scanning to form an image of the burn area. We here present a prototype for a hybrid DCS and LSI technique called speckle contrast diffuse correlation spectroscopy (scDCS) with the novel approach of using a laser line as a source and using the speckle contrast of averaged images to obtain an estimate of static scattering in the tissue. This will allow for fast non-contact 1D scanning to perform 3D tomographic imaging, making quantitative estimates of the depth and area of the coagulation zone from burn wounds. Simulations and experimental results from a volumetric flow phantom and a gelatin wedge phantom show promise to determine coagulation depth. The aim is to develop a method that, in the future, could provide more quantitative estimates of coagulation depth in partial thickness burn wounds to better estimate when surgery is needed.
部分厚度烧伤的创面部分穿过真皮层,使许多痛觉感受器完好无损,从而使创面非常疼痛。由于疼痛难忍,快速评估烧伤深度非常重要,以免耽误必要的伤口手术。皮肤血流激光斑点成像(LSI)有助于发现血流受损的严重凝固区。不过,LSI 测量通常过于浅表,无法正确触及成人真皮层的全部深度,也无法分辨深度血流。漫射相关光谱(DCS)使用不同的源-检测器分离来区分血流深度,但需要耗时的二维扫描来形成烧伤区域的图像。我们在此介绍一种混合 DCS 和 LSI 技术的原型,称为斑点对比度漫反射相关光谱(scDCS),其新颖之处在于使用激光线作为光源,并利用平均图像的斑点对比度来估算组织中的静态散射。这样就可以通过快速非接触式一维扫描进行三维断层成像,对烧伤创面凝固区的深度和面积进行定量估计。体积流动模型和明胶楔形模型的模拟和实验结果表明,有望确定凝固深度。我们的目标是开发一种方法,将来可以对部分厚度烧伤创面的凝固深度进行更定量的估计,以便更好地估计何时需要手术。
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引用次数: 0
Monitoring of neoadjuvant chemotherapy through time domain diffuse optics: breast tissue composition changes and collagen discriminative potential 通过时域漫反射光学监控新辅助化疗:乳腺组织成分变化和胶原蛋白鉴别潜力
IF 3.4 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-27 DOI: 10.1364/boe.527968
Nikhitha Mule, Giulia Maffeis, Rinaldo Cubeddu, Carolina Santangelo, Giampaolo Bianchini, Pietro Panizza, Paola Taroni
The purpose of this clinical study is to test a broad spectral range (635-1060 nm) time-domain diffuse optical spectroscopy in monitoring the response of breast cancer patients to neoadjuvant chemotherapy (NAC). The broadband operation allows us to fully analyze tissue composition in terms of hemoglobin, water, lipids and collagen concentration, which has never been systematically studied until now during the course of therapy. Patients are subjected to multiple breast optical imaging sessions, each one performed at different stages of NAC, both on tumor-bearing and contralateral healthy breasts. We correlate the optical results with conventional imaging techniques and pathological response. Preliminary outcomes on 10 patients’ data show an average significant reduction in the concentrations of oxy-hemoglobin (-53%, p = 0.0020), collagen (-36%, p = 0.0039) and water (-15%, p = 0.0195), and increase in lipids (+39%, p = 0.0137) from baseline to the end of therapy in the tumor-bearing breast of patients who responded to therapy at least partially. With respect to scattering, the scattering amplitude, a, increases slightly (+15%, p = 0.0039) by the end of the therapy compared to the baseline, while the scattering slope, b, shows no significant change (+4%, p = 0.9219). Some change in the constituents’ concentrations was also noticed in the contralateral healthy breast, even though it was significant only for oxy-hemoglobin concentration. We observed that collagen seems to be the only component distinguishing between complete and partial responders by the end of 2-3 weeks from the baseline. In the complete responder group, collagen significantly decreased after 2-3 weeks with respect to baseline (p = 0.0423). While the partial responder group also showed a decrease, it did not reach statistical significance (p = 0.1012). This suggests that collagen could serve as a potential biomarker to measure NAC effectiveness early during treatment. Even though obtained on a small group of patients, these initial results are consistent with those of standard medical modalities and highlight the sensitivity of the technique to changes that occur in breast composition during NAC.
这项临床研究的目的是测试宽光谱范围(635-1060 nm)时域漫射光学光谱在监测乳腺癌患者对新辅助化疗(NAC)反应方面的应用。宽带操作使我们能够全面分析血红蛋白、水、脂质和胶原蛋白浓度方面的组织成分,迄今为止,我们从未在治疗过程中对这些成分进行过系统研究。我们对患者进行了多次乳腺光学成像,每次成像都是在新农合的不同阶段对肿瘤患者和对侧健康乳房进行的。我们将光学成像结果与传统成像技术和病理反应相关联。10 名患者数据的初步结果显示,从基线到治疗结束,至少部分对治疗有反应的患者的肿瘤乳房中氧合血红蛋白(-53%,p = 0.0020)、胶原蛋白(-36%,p = 0.0039)和水(-15%,p = 0.0195)的浓度平均显著降低,而脂质(+39%,p = 0.0137)则有所增加。在散射方面,与基线相比,散射振幅a在治疗结束时略有增加(+15%,p = 0.0039),而散射斜率b则无明显变化(+4%,p = 0.9219)。对侧健康乳房中的成分浓度也发生了一些变化,但只有氧合血红蛋白浓度发生了显著变化。我们观察到,从基线算起,2-3 周后,胶原蛋白似乎是区分完全应答者和部分应答者的唯一成分。在完全应答组中,2-3 周后胶原蛋白与基线相比明显减少(p = 0.0423)。虽然部分反应者组也出现了下降,但未达到统计学意义(p = 0.1012)。这表明,胶原蛋白可作为一种潜在的生物标志物,在治疗早期衡量 NAC 的有效性。尽管这只是在一小部分患者身上获得的结果,但这些初步结果与标准医疗模式的结果一致,并凸显了该技术对 NAC 期间乳房成分变化的敏感性。
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引用次数: 0
Learning-based distortion correction enables proximal-scanning endoscopic OCT elastography. 基于学习的失真校正实现了近端扫描内窥镜 OCT 弹性成像。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-26 eCollection Date: 2024-07-01 DOI: 10.1364/BOE.528522
Haoran Zhang, Chengfu Gu, Qi Lan, Weiyi Zhang, Chang Liu, Jianlong Yang

Proximal rotary scanning is predominantly used in the clinical practice of endoscopic and intravascular OCT, mainly because of the much lower manufacturing cost of the probe compared to distal scanning. However, proximal scanning causes severe beam stability issues (also known as non-uniform rotational distortion, NURD), which hinders the extension of its applications to functional imaging, such as OCT elastography (OCE). In this work, we demonstrate the abilities of learning-based NURD correction methods to enable the imaging stability required for intensity-based OCE. Compared with the previous learning-based NURD correction methods that use pseudo distortion vectors for model training, we propose a method to extract real distortion vectors from a specific endoscopic OCT system, and validate its superiority in accuracy under both convolutional-neural-network- and transformer-based learning architectures. We further verify its effectiveness in elastography calculations (digital image correlation and optical flow) and the advantages of our method over other NURD correction methods. Using the air pressure of a balloon catheter as a mechanical stimulus, our proximal-scanning endoscopic OCE could effectively differentiate between areas of varying stiffness of atherosclerotic vascular phantoms. Compared with the existing endoscopic OCE methods that measure only in the radial direction, our method could achieve 2D displacement/strain distribution in both radial and circumferential directions.

近端旋转扫描主要用于内窥镜和血管内 OCT 的临床实践,这主要是因为与远端扫描相比,探头的制造成本要低得多。然而,近端扫描会导致严重的光束稳定性问题(也称为非均匀旋转变形,NURD),这阻碍了其在功能成像(如 OCT 弹性成像,OCE)方面的应用。在这项工作中,我们展示了基于学习的 NURD 校正方法实现基于强度的 OCE 所需的成像稳定性的能力。与之前使用伪失真向量进行模型训练的基于学习的 NURD 校正方法相比,我们提出了一种从特定内窥镜 OCT 系统中提取真实失真向量的方法,并验证了其在基于卷积神经网络和转换器的学习架构下的准确性优势。我们进一步验证了它在弹性成像计算(数字图像相关性和光流)中的有效性,以及我们的方法与其他 NURD 校正方法相比的优势。利用球囊导管的气压作为机械刺激,我们的近端扫描内窥镜 OCE 可以有效区分动脉粥样硬化血管模型的不同僵硬度区域。与仅在径向进行测量的现有内窥镜 OCE 方法相比,我们的方法可在径向和周向实现二维位移/应变分布。
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引用次数: 0
Time-domain diffuse correlation spectroscopy at large source detector separation for cerebral blood flow recovery. 用于脑血流恢复的大源探测器分离时域漫反射相关光谱。
IF 2.9 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-26 eCollection Date: 2024-07-01 DOI: 10.1364/BOE.523514
Neda Mogharari, Stanisław Wojtkiewicz, Dawid Borycki, Adam Liebert, Michał Kacprzak

Time-domain diffuse correlation spectroscopy (td-DCS) enables the depth discrimination in tissue's blood flow recovery, considering the fraction of photons detected with higher time of flight (TOF) and longer pathlength through the tissue. However, the recovery result depends on factors such as the instrument response function (IRF), analyzed TOF gate start time, gate width and the source-detector separation (SDS). In this research we evaluate the performance of the td-DCS technique at three SDSs of 1.5, 2 and 2.5 cm to recover cerebral blood flow (CBF). To do that we presented comprehensive characterization of the td-DCS system through a series of phantom experiments. First by quality metrices such as coefficient of variation and contrast-to-noise ratios, we identified optimal time gate(s) of the TOF to extract dynamics of particles. Then using sensitivity metrices, each SDS ability to detect dynamics of particles in superficial and deeper layer was evaluated. Finally, td-DCS at each SDS was tested on healthy volunteers during cuff occlusion test and breathing tasks. According to phantom measurements, the sensitivity to estimate perfusion within the deep layer located at depth of 1.5 cm from the surface can be increased more than two times when the SDS increases from 1.5 cm to 2.5 cm.

时域漫反射相关光谱(td-DCS)能够在组织血流恢复中进行深度分辨,考虑的是以更高的飞行时间(TOF)和更长的组织路径长度检测到的光子分数。然而,恢复结果取决于各种因素,如仪器响应函数(IRF)、分析 TOF 栅极启动时间、栅极宽度和源-探测器分离(SDS)。在这项研究中,我们评估了td-DCS 技术在 1.5、2 和 2.5 厘米三个 SDS 下恢复脑血流(CBF)的性能。为此,我们通过一系列假体实验对td-DCS系统进行了全面鉴定。首先,通过变异系数和对比-噪声比等质量指标,我们确定了提取颗粒动态的 TOF 最佳时间门。然后,使用灵敏度指标评估了每种 SDS 检测浅层和深层粒子动态的能力。最后,在袖带闭塞测试和呼吸任务中,对健康志愿者进行了每个 SDS 的 td-DCS 测试。根据模型测量结果,当 SDS 从 1.5 厘米增加到 2.5 厘米时,对位于距离表面 1.5 厘米深处的深层灌注的估计灵敏度可提高两倍以上。
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引用次数: 0
Reducing manual labeling requirements and improved retinal ganglion cell identification in 3D AO-OCT volumes using semi-supervised learning 利用半监督学习减少人工标记要求并改进三维 AO-OCT 图像中视网膜神经节细胞的识别
IF 3.4 2区 医学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-06-26 DOI: 10.1364/boe.526053
Mengxi Zhou, Yue Zhang, Amin Karimi Monsefi, Stacey S. Choi, Nathan Doble, Srinivasan Parthasarathy, Rajiv Ramnath
Adaptive optics-optical coherence tomography (AO-OCT) allows for the three-dimensional visualization of retinal ganglion cells (RGCs) in the living human eye. Quantitative analyses of RGCs have significant potential for improving the diagnosis and monitoring of diseases such as glaucoma. Recent advances in machine learning (ML) have made possible the automatic identification and analysis of RGCs within the complex three-dimensional retinal volumes obtained with such imaging. However, the current state-of-the-art ML approach relies on fully supervised training, which demands large amounts of training labels. Each volume requires many hours of expert manual annotation. Here, two semi-supervised training schemes are introduced, (i) cross-consistency training and (ii) cross pseudo supervision that utilize unlabeled AO-OCT volumes together with a minimal set of labels, vastly reducing the labeling demands. Moreover, these methods outperformed their fully supervised counterpart and achieved accuracy comparable to that of human experts.
自适应光学-光学相干断层扫描(AO-OCT)可实现活体人眼视网膜神经节细胞(RGC)的三维可视化。对 RGCs 的定量分析在改善青光眼等疾病的诊断和监测方面具有巨大潜力。机器学习(ML)的最新进展使得在这种成像技术获得的复杂三维视网膜体积中自动识别和分析 RGC 成为可能。然而,目前最先进的 ML 方法依赖于完全监督训练,这需要大量的训练标签。每个体积都需要专家人工标注许多小时。本文介绍了两种半监督训练方案:(i) 交叉一致性训练和 (ii) 交叉伪监督,它们利用未标记的 AO-OCT 容量和最小标签集,大大降低了标记需求。此外,这些方法的效果优于完全监督的方法,其准确度可与人类专家相媲美。
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引用次数: 0
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Biomedical optics express
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