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Fluorescence Spectroscopy With Temperature Functional Tests in the Assessment of Markers of Intracellular Energy Metabolism: Spatial Heterogeneity and Reproducibility of Measurements 荧光光谱与温度功能测试在细胞内能量代谢标志物评估中的应用:测量的空间异质性和可重复性。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-28 DOI: 10.1002/jbio.202400294
Ekaterina Ryzhkova, Tatyana Morgunova, Elena Potapova, Ivan Ryzhkov, Valentin Fadeyev

The fluorescence intensities of the cellular respiratory cofactors NADH (reduced nicotinamide adenine dinucleotide) and FAD++ (oxidized flavin adenine dinucleotide) reflect energy metabolism in skin and other tissues and can be quantified in vivo by fluorescence spectroscopy (FS). However, the variability of physiological parameters largely determines the reproducibility of measurement results and the reliability of the diagnostic test. In this prospective study, we evaluated the interday reproducibility of NADH and FAD++ fluorescence intensity measurements in the skin of 51 healthy volunteers assessed by the FS at baseline, after local cooling (10°C) and heating of the skin (35°C). Results showed that the fluorescence amplitude of NADH (AFNADH) in forearm skin was the most reproducible of the FS parameters studied. Assessment of AFNADH in the dorsal forearm in combination with a thermal functional test is the most promising method for clinical use for assessing energy metabolism in the skin.

细胞呼吸辅助因子 NADH(还原型烟酰胺腺嘌呤二核苷酸)和 FAD++(氧化型黄素腺嘌呤二核苷酸)的荧光强度反映了皮肤和其他组织的能量代谢情况,可通过荧光光谱法(FS)进行活体量化。然而,生理参数的可变性在很大程度上决定了测量结果的可重复性和诊断测试的可靠性。在这项前瞻性研究中,我们评估了 51 名健康志愿者皮肤中 NADH 和 FAD++ 荧光强度测量值在基线、局部冷却(10°C)和皮肤加热(35°C)后的重现性。结果显示,在所研究的 FS 参数中,前臂皮肤中 NADH(AFNADH)荧光振幅的重现性最好。结合热功能测试评估前臂背侧的 AFNADH 是最有希望用于临床评估皮肤能量代谢的方法。
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引用次数: 0
Downconversion Master Slave OCT With a Bidirectional Sweeping Laser 使用双向扫描激光器的下变频主从 OCT
IF 2.8 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-28 DOI: 10.1002/jbio.202400201
A. Martinez Jimenez, R. Cernat, A. Bradu, R. Riha, E. A. Proano Grijalva, B. O. Meyer, T. Ansbaek, K. Yvind, A. Podoleanu
This paper explores the challenges of signal processing when using optical coherence tomography (OCT) imaging instruments driven by asymmetric MHz bidirectional sweeping lasers. A downconversion master–slave (DMS) method is proposed as a viable alternative to the traditional OCT protocol. Unlike conventional swept source OCT, which requires a separate calibration for each sweep, the DMS approach does not require calibration of the acquired channeled spectra; its operation is independent of the tuning direction. We demonstrate the practicality of the DMS method with en‐face OCT images obtained with an OCT instrument equipped with a fast bidirectional swept laser (tuning speed 1.6 MHz) and a slow acquisition card of only 2.5 MS/s sampling rate.
本文探讨了使用非对称 MHz 双向扫描激光器驱动的光学相干断层扫描(OCT)成像仪器时信号处理所面临的挑战。本文提出了一种下转换主从(DMS)方法,作为传统 OCT 协议的可行替代方案。传统的扫描光源 OCT 需要对每次扫描进行单独校准,而 DMS 方法则不同,它不需要对获取的通道光谱进行校准;其操作与调谐方向无关。我们用配备了快速双向扫频激光器(调谐速度为 1.6 MHz)和采样率仅为 2.5 MS/s 的慢速采集卡的 OCT 仪器获得的面内 OCT 图像证明了 DMS 方法的实用性。
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引用次数: 0
Motion Artifact Correction for OCT Microvascular Images Based on Image Feature Matching 基于图像特征匹配的 OCT 微血管图像运动伪影校正。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-28 DOI: 10.1002/jbio.202400198
Xudong Chen, Zongqing Ma, Chongyang Wang, Jiaqi Cui, Fan Fan, Xinxiao Gao, Jiang Zhu

Optical coherence tomography angiography (OCTA), a functional extension of optical coherence tomography (OCT), is widely employed for high-resolution imaging of microvascular networks. However, due to the relatively low scan rate of OCT, the artifacts caused by the involuntary bulk motion of tissues severely impact the visualization of microvascular networks. This study proposes a fast motion correction method based on image feature matching for OCT microvascular images. First, the rigid motion-related mismatch between B-scans is compensated through the image feature matching based on the improved oriented FAST and rotated BRIEF algorithm. Then, the axial motion within A-scan lines in each B-scan image is corrected according to the displacement deviation between the detected boundaries achieved by the Scharr operator in a non-rigid transformation manner. Finally, an optimized intensity-based Doppler variance algorithm is developed to enhance the robustness of the OCTA imaging. The experimental results demonstrate the effectiveness of the method.

光学相干断层血管成像(OCTA)是光学相干断层成像(OCT)的功能扩展,被广泛用于微血管网络的高分辨率成像。然而,由于 OCT 的扫描速率相对较低,组织的不自主运动造成的伪影严重影响了微血管网络的可视化。本研究提出了一种基于图像特征匹配的 OCT 微血管图像快速运动校正方法。首先,基于改进的定向 FAST 和旋转 BRIEF 算法,通过图像特征匹配补偿 B 扫描之间与刚性运动相关的不匹配。然后,根据 Scharr 算子以非刚性变换方式实现的检测边界之间的位移偏差,校正每个 B 扫描图像中 A 扫描线内的轴向运动。最后,开发了一种基于强度的多普勒方差优化算法,以增强 OCTA 成像的鲁棒性。实验结果证明了该方法的有效性。
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引用次数: 0
Dual-Modal Optical Imaging of Tissue Perfusion in Response to Cooling Stimulation Facilitates Early Detection of Pressure Ulcer 组织灌注对冷却刺激响应的双模式光学成像有助于早期发现褥疮。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-27 DOI: 10.1002/jbio.202400188
Qingdong Zhang, Peng Liu, Pengfei Shao, Mingzhai Sun, Peng Yao, Shuwei Shen, Yang Zhang, Ming Wu, Ronald X. Xu

Pressure ulcers present a significant human and economic challenge, lacking a reliable method for early detection. To address this, we developed a system capable of early detection by using cooling stimulation and dynamic data acquisition techniques to monitor blood perfusion and skin temperature. The system consists of laser speckle perfusion imaging and thermal imaging. And we performed simulations to demonstrate that the system is capable of detect tissue damage across multiple layers, from superficial to deep. Testing on a rabbit ear model demonstrated that this approach, which combines dynamic perfusion and temperature parameters, effectively distinguishes early pressure ulcer areas from normal skin with a significant p value of 0.0015. This distinction was more precise compared to methods relying solely on static parameters or one parameter. Our study thereby offers a promising advancement in the proactive management and prevention of pressure ulcers.

由于缺乏早期检测的可靠方法,褥疮给人类和经济带来了巨大挑战。为了解决这个问题,我们开发了一种能够利用冷却刺激和动态数据采集技术监测血液灌注和皮肤温度的早期检测系统。该系统由激光斑点灌注成像和热成像组成。我们进行了模拟实验,证明该系统能够检测从浅层到深层的多层组织损伤。在兔耳模型上进行的测试表明,这种方法结合了动态灌注和温度参数,能有效区分早期压疮区域和正常皮肤,其显著的 p 值为 0.0015。与仅依靠静态参数或一种参数的方法相比,这种区分更为精确。因此,我们的研究为压疮的主动管理和预防带来了希望。
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引用次数: 0
Effect of photobiomodulation in an experimental in vitro model of asthma-Copd overlap 光生物调节在哮喘-镉重叠实验体外模型中的效果。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-12 DOI: 10.1002/jbio.202400124
Marina Bertoni Guerra, Kelly Gomes Santana, Marcos Momolli, Rodrigo Labat, Maria Cristina Chavantes, Stella Regina Zammuner, José Antonio Silva Júnior, Renata Kelly da Palma, Flavio Aimbire, Ana Paula Ligeiro de Oliveira

The objective of the study was to evaluate the effect of photobiomodulation (PBM) with laser on the inflammatory process in an experimental in vitro model of ACO. The groups were: (1) human bronchial epithelial cells (BEAS-2B); (2) BEAS-2B cells treated with dexamethasone; (3) BEAS-2B cells irradiated with laser; (4) BEAS-2B cells stimulated with cigarette smoke extract (CSE) + House Dust Mite (HDM); (5) BEAS-2B cells stimulated with CSE + HDM and treated with dexamethasone; (6) BEAS-2B cells incubated with CSE + HDM and irradiated with laser. After 24 h, cytokines were quantified. There was a reduction in TNF-α, IL-1β, IL-6, IL-4, IL-5, IL-13, IL-17, IL-21, IL-23, and an increase in IL-10 and IFN-γ in cells from the laser-irradiated ACO group compared to only ACO group. With these results, we can suggest that photobiomodulation acts in the modulation of inflammation observed in ACO, and may be a treatment option.

该研究旨在评估激光光生物调节(PBM)对 ACO 实验性体外模型炎症过程的影响。实验组包括(1) 人支气管上皮细胞(BEAS-2B);(2) 用地塞米松处理的 BEAS-2B 细胞;(3) 用激光照射的 BEAS-2B 细胞;(4) 用香烟烟雾提取物(CSE)+屋尘螨(HDM)刺激的 BEAS-2B 细胞;(5)用 CSE + HDM 刺激 BEAS-2B 细胞并用地塞米松处理;(6)用 CSE + HDM 培养 BEAS-2B 细胞并用激光照射。24 小时后,对细胞因子进行量化。与只用 ACO 组相比,用激光照射 ACO 组的细胞中 TNF-α、IL-1β、IL-6、IL-4、IL-5、IL-13、IL-17、IL-21、IL-23 减少,IL-10 和 IFN-γ 增加。根据这些结果,我们可以认为,光生物调制对 ACO 中观察到的炎症有调节作用,可能是一种治疗选择。
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引用次数: 0
Signatures of top versus bottom illuminations and their predicted implications for infrared transmission microspectroscopy 顶部照明与底部照明的特征及其对红外透射显微光谱学的影响预测。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-11 DOI: 10.1002/jbio.202400079
Beibei Kong, Reinhold Blümel, Pasi Ylä-Oijala, Henrik Wallén, Ari Sihvola, Achim Kohler

Since both top and bottom illuminations are widely used in infrared transmission measurements, in this paper, we study the effects of different illuminations on the signatures in infrared microspectroscopy. By simulating a series of dielectric samples, we show that their extinction efficiency, Qext, remains unchanged when the direction of the incident plane wave is reversed, even though the field distributions both inside and outside of the sample may be dramatically different. We find features in Qext that are correlated with whispering gallery modes for one beam direction and correspond to completely different field distributions for the opposite beam direction. In addition, by linking the optical theorem and the reciprocity relation of far-field scattered field, we rigorously prove the invariance of Qext for arbitrary dielectric targets under opposite plane-wave illuminations. Furthermore, we show the difference in the apparent absorbance spectrum for opposite beam directions when considering numerical apertures.

由于顶部和底部照明在红外透射测量中都被广泛使用,本文研究了不同照明对红外显微光谱分析中信号的影响。通过模拟一系列电介质样品,我们发现它们的消光效率 Q ext $$ {Q}_{mathrm{ext}}$$$ ,当入射平面波的方向相反时保持不变,即使样品内外的场分布可能有很大不同。我们在 Q ext $$ {Q}_{mathrm{ext}} 中发现了与耳语相关的特征。$$ 中的特征,这些特征在一个光束方向上与耳语画廊模式相关,而在相反的光束方向上则与完全不同的场分布相对应。此外,通过将光学定理与远场散射场的互易关系联系起来,我们严格证明了 Q ext $$ {Q}_{mathrm{ext}} 的不变性。$$ 在相反平面波照射下对任意介质目标的不变性。此外,我们还展示了在考虑数值孔径时,相反光束方向的视吸收光谱的差异。
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引用次数: 0
Texture-based speciation of otitis media-related bacterial biofilms from optical coherence tomography images using supervised classification 利用监督分类法从光学相干断层扫描图像中对中耳炎相关细菌生物膜进行基于纹理的分类。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-05 DOI: 10.1002/jbio.202400075
Farzana R. Zaki, Guillermo L. Monroy, Jindou Shi, Kavya Sudhir, Stephen A. Boppart

Otitis media (OM), a highly prevalent inflammatory middle-ear disease in children worldwide, is commonly caused by an infection, and can lead to antibiotic-resistant bacterial biofilms in recurrent/chronic OM cases. A biofilm related to OM typically contains one or multiple bacterial species. OCT has been used clinically to visualize the presence of bacterial biofilms in the middle ear. This study used OCT to compare microstructural image texture features from bacterial biofilms. The proposed method applied supervised machine-learning-based frameworks (SVM, random forest, and XGBoost) to classify multiple species bacterial biofilms from in vitro cultures and clinically-obtained in vivo images from human subjects. Our findings show that optimized SVM-RBF and XGBoost classifiers achieved more than 95% of AUC, detecting each biofilm class. These results demonstrate the potential for differentiating OM-causing bacterial biofilms through texture analysis of OCT images and a machine-learning framework, offering valuable insights for real-time in vivo characterization of ear infections.

中耳炎(OM)是全球儿童高发的一种中耳炎性疾病,通常由感染引起,在复发性/慢性中耳炎病例中可导致抗生素耐药细菌生物膜。与 OM 相关的生物膜通常包含一种或多种细菌。OCT 已被临床用于观察中耳是否存在细菌生物膜。本研究使用 OCT 比较细菌生物膜的微结构图像纹理特征。所提出的方法应用了基于机器学习的监督框架(SVM、随机森林和 XGBoost),对体外培养和临床获得的人体活体图像中的多种细菌生物膜进行分类。我们的研究结果表明,经过优化的 SVM-RBF 和 XGBoost 分类器的 AUC 超过了 95%,能检测出每一类生物膜。这些结果证明了通过对 OCT 图像的纹理分析和机器学习框架来区分 OM 致病细菌生物膜的潜力,为耳部感染的实时活体特征描述提供了有价值的见解。
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引用次数: 0
Multi-color two-laser super-resolution structured illumination microscopy for the visualization of multi-organelle in living cells 用于活细胞多细胞器可视化的多色双激光超分辨率结构照明显微镜。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-04 DOI: 10.1002/jbio.202400154
Xuejuan Hu, Yadan Tan, Yujie Huang, Jianze Ye, Yifei Liang, Xiaokun Yang, Hengliang Wang, Zihao Cheng, Lihu Wang, Shiqian Liu, Minfei Li, Zhengdi He, Qianding Gao, Jingli Zhong

In this study, we introduced a novel dual-laser multi-color imaging system. Integrated with a multi-channel filter wheel, this system compared three spectral decontamination algorithms (nonnegative matrix factorization [NMF], RCAN, and PICASSO) showcasing its efficacy in achieving four-color imaging with only two laser sources. Combined with a reliable image reconstruction algorithm, the spatial resolution of four channels super-resolution four-color images reached 130, 125, 133, and 132 nm, respectively. Lipid droplets, mitochondria, lysosomes, and nuclei from the mouse hepatocytes (AML12), human neuroblastoma cells (SH-SY5Y), mouse hippocampal neuronal cells (HT-22), and immortalized murine bone marrow-derived macrophages were imaged. At the same time, the chromatin condensation, nuclear contraction, DNA fragmentation, apoptotic body formation, as well as the fusion of Mito and Lyso involved in mitochondrial autophagy were observed in HT-22 and SH-SY5Y cells suffering oxidative stress. Our multi-color SIM imaging system establishes a powerful platform for dynamic organelle studies and other high-resolution investigations in live cells.

在这项研究中,我们介绍了一种新型双激光多色成像系统。该系统集成了一个多通道滤光轮,比较了三种光谱净化算法(非负矩阵因式分解[NMF]、RCAN 和 PICASSO),展示了其在仅使用两个激光源实现四色成像方面的功效。结合可靠的图像重建算法,四通道超分辨率四色图像的空间分辨率分别达到了 130、125、133 和 132 nm。对小鼠肝细胞(AML12)、人神经母细胞瘤细胞(SH-SY5Y)、小鼠海马神经元细胞(HT-22)和永生化小鼠骨髓巨噬细胞的脂滴、线粒体、溶酶体和细胞核进行了成像。同时,在遭受氧化应激的 HT-22 和 SH-SY5Y 细胞中观察到染色质凝结、核收缩、DNA 断裂、凋亡体形成以及参与线粒体自噬的 Mito 和 Lyso 融合。我们的多色 SIM 成像系统为活细胞中的动态细胞器研究和其他高分辨率研究建立了一个强大的平台。
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引用次数: 0
Intelligent skin-removal photoacoustic computed tomography for human based on deep learning 基于深度学习的智能皮肤去除光声计算机断层扫描。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-08-02 DOI: 10.1002/jbio.202400197
Ning Wang, Tao Chen, Chengbo Liu, Jing Meng

Photoacoustic computed tomography (PACT) has centimeter-level imaging ability and can be used to detect the human body. However, strong photoacoustic signals from skin cover deep tissue information, hindering the frontal display and analysis of photoacoustic images of deep regions of interest. Therefore, we propose a 2.5 D deep learning model based on feature pyramid structure and single-type skin annotation to extract the skin region, and design a mask generation algorithm to remove skin automatically. PACT imaging experiments on the human periphery blood vessel verified the correctness our proposed skin-removal method. Compared with previous studies, our method exhibits high robustness to the uneven illumination, irregular skin boundary, and reconstruction artifacts in the images, and the reconstruction errors of PACT images decreased by 20% ~ 90% with a 1.65 dB improvement in the signal-to-noise ratio at the same time. This study may provide a promising way for high-definition PACT imaging of deep tissues.

光声计算机断层扫描(PACT)具有厘米级成像能力,可用于探测人体。然而,来自皮肤的强光声信号会覆盖深层组织信息,阻碍深层感兴趣区域光声图像的正面显示和分析。因此,我们提出了一种基于特征金字塔结构和单一类型皮肤标注的 2.5 D 深度学习模型来提取皮肤区域,并设计了一种自动去除皮肤的掩膜生成算法。对人体外周血管的 PACT 成像实验验证了我们提出的皮肤去除方法的正确性。与之前的研究相比,我们的方法对图像中不均匀光照、不规则皮肤边界和重建伪影表现出很高的鲁棒性,PACT 图像的重建误差降低了 20% ~ 90%,信噪比同时提高了 1.65 dB。这项研究为深部组织的高清 PACT 成像提供了一条可行的途径。
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引用次数: 0
Angiogenesis-elicited spectral responses of early invasive skin melanoma: Implications for the evaluation of lesion progression 早期浸润性皮肤黑色素瘤的血管生成诱导光谱反应:对评估病变进展的意义。
IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2024-07-30 DOI: 10.1002/jbio.202400208
Gladimir V. G. Baranoski, Petri M. Varsa

Early invasive skin melanoma (EISM) associated with partial tumor invasion to the thin and optically complex papillary dermis (PD) represents a critical stage before the onset of metastasis. EISM lesions may be accompanied by angiogenesis, which can alter the PD's blood and fibril contents. A comprehensive understanding about these interconnected processes is essential for enhancing the efficacy of EISM optical evaluation methodologies. Employing a first-principles computational approach supported by measured data, we systematically assess the impact that angiogenesis can have on the EISM's spectral responses. Our findings indicate that these responses are discernibly affected by angiogenesis under distinct physiological conditions, with more substantial tissue alterations leading to accentuated spectral changes in the 550–600 nm region. Accordingly, we propose the use of a customized low-cost spectral index to monitor these processes. Furthermore, our investigation provides a high-fidelity in silico platform for interdisciplinary research on the photobiology of evolving skin melanomas.

早期浸润性皮肤黑色素瘤(EISM)伴有肿瘤部分侵入薄而光学结构复杂的真皮乳头层(PD),是转移开始前的一个关键阶段。EISM病变可能伴有血管生成,这会改变真皮乳头的血液和纤维内容。全面了解这些相互关联的过程对于提高 EISM 光学评估方法的有效性至关重要。在测量数据的支持下,我们采用第一原理计算方法,系统地评估了血管生成对 EISM 光谱响应的影响。我们的研究结果表明,在不同的生理条件下,血管生成会对这些响应产生明显的影响,组织的实质性改变会导致 550-600 纳米区域的光谱变化加剧。因此,我们建议使用定制的低成本光谱指数来监测这些过程。此外,我们的研究还为不断演变的皮肤黑色素瘤的光生物学跨学科研究提供了一个高保真硅学平台。
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引用次数: 0
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Journal of Biophotonics
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