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Cellphone-based hand-held microplate reader for point-of-care ELISA testing (Conference Presentation) 基于手机的手持式酶联免疫吸附试验微孔板阅读器(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2209125
B. Berg, Bingen Cortazar, Derek K. Tseng, Haydar Ozkan, S. Feng, Qingshan Wei, R. Y. Chan, J. Burbano, Qamar Farooqui, M. Lewinski, D. Di Carlo, O. Garner, A. Ozcan
Enzyme-linked immunosorbent assay (ELISA) in a microplate format has been a gold standard first-line clinical test for diagnosis of various diseases including infectious diseases. However, this technology requires a relatively large and expensive multi-well scanning spectrophotometer to read and quantify the signal from each well, hindering its implementation in resource-limited-settings. Here, we demonstrate a cost-effective and handheld smartphone-based colorimetric microplate reader for rapid digitization and quantification of immunoserology-related ELISA tests in a conventional 96-well plate format at the point of care (POC). This device consists of a bundle of 96 optical fibers to collect the transmitted light from each well of the microplate and direct all the transmission signals from the wells onto the camera of the mobile-phone. Captured images are then transmitted to a remote server through a custom-designed app, and both quantitative and qualitative diagnostic results are returned back to the user within ~1 minute per 96-well plate by using a machine learning algorithm. We tested this mobile-phone based micro-plate reader in a clinical microbiology lab using FDA-approved mumps IgG, measles IgG, and herpes simplex virus IgG (HSV-1 and HSV-2) ELISA tests on 1138 remnant patient samples (roughly 50% training and 50% testing), and achieved an overall accuracy of ~99% or higher for each ELISA test. This handheld and cost-effective platform could be immediately useful for large-scale vaccination monitoring in low-infrastructure settings, and also for other high-throughput disease screening applications at POC.
酶联免疫吸附试验(ELISA)已成为包括传染病在内的各种疾病诊断的一线临床检测的金标准。然而,该技术需要一个相对较大且昂贵的多井扫描分光光度计来读取和量化每口井的信号,这阻碍了其在资源有限的环境下的实施。在这里,我们展示了一种具有成本效益的手持式智能手机比色微孔板阅读器,用于在护理点(POC)以传统的96孔板格式快速数字化和定量免疫血清学相关ELISA测试。该装置由一束96根光纤组成,用于收集微孔板每孔的透射光,并将所有传输信号引导到手机的摄像头上。然后通过定制的应用程序将捕获的图像传输到远程服务器,并使用机器学习算法在每96孔板约1分钟内将定量和定性诊断结果返回给用户。我们在临床微生物学实验室使用fda批准的腮腺炎IgG、麻疹IgG和单纯疱疹病毒IgG (HSV-1和HSV-2) ELISA检测1138例剩余患者样本(大约50%训练和50%检测),对这种基于手机的微板读取器进行了测试,每次ELISA检测的总体准确率达到~99%或更高。这种具有成本效益的手持式平台可立即用于低基础设施环境中的大规模疫苗接种监测,也可用于POC的其他高通量疾病筛查应用。
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引用次数: 0
Morphological study of human sweat ducts for the investigation of THz-wave interaction (Conference Presentation) 人体汗腺形态学研究用于太赫兹波相互作用的研究(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2211618
K. Kawase, S. Tripathi
Recently, some studies reported that the sweat ducts act as a low-Q-factor helical antenna due to their helical structure, and resonate in the terahertz frequency range according to their structural parameters. According to the antenna theory, when the duct works as a helical antenna, the dimension of the helix plays a key role to determine the frequency of resonance. Therefore, the accurate determination of structural parameters of sweat duct is crucially important to obtain the reliable frequency of resonance and modes of operations. Therefore, here we performed the optical coherence tomography (OCT) of human subjects on their palm and foot to investigate the density, distribution and morphological features of sweat ducts. Moreover, we measured the dielectric properties of stratum corneum using terahertz time domain spectroscopy and based upon this information, we determined the frequency of resonance. We recruited 32 subjects for the measurement and the average duct diameter was 95±11μm. Based upon this information on diameter of duct and THz dielectric properties of stratum corneum (ε=5.1±1.3), we have calculated the frequency of resonance of sweat duct. Finally, we determined that the center frequency of resonance was 442±76 GHz. We believe that these findings will facilitate further investigation of the THz-skin interaction and provide guidelines for safety levels with respect to human exposure. We will also report on the EEG measurement while being shined by micro watt order THz waves.
近年来,有研究报道汗腺由于其螺旋结构,可以作为低q因子的螺旋天线,根据其结构参数在太赫兹频率范围内产生共振。根据天线理论,当风管作为螺旋天线工作时,螺旋的尺寸对决定谐振频率起着关键作用。因此,准确确定汗道的结构参数对于获得可靠的谐振频率和工作方式至关重要。因此,本研究对人体手掌和足部进行光学相干断层扫描(OCT),研究汗腺的密度、分布和形态特征。此外,我们还利用太赫兹时域光谱测量了角质层的介电特性,并在此基础上确定了共振频率。我们招募了32名受试者进行测量,平均导管直径为95±11μm。根据导管直径和角质层的太赫兹介电特性(ε=5.1±1.3),计算出了汗液导管的谐振频率。最后,我们确定了共振的中心频率为442±76 GHz。我们相信这些发现将有助于进一步研究太赫兹与皮肤的相互作用,并为人类暴露的安全水平提供指导。我们还将报道在微瓦级太赫兹波照射下的脑电图测量。
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引用次数: 0
Preanalytical considerations in detection of colorectal cancer in blood serum using Raman molecular imaging (Conference Presentation) 使用拉曼分子成像检测血清中大肠癌的分析前考虑(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2212776
P. Treado, Shona Stewart, Aaron Smith, H. Kirschner, C. Post, B. Overholt
Colorectal cancer (CRC) is the third most common cancer in men and women in the United States. Raman Molecular Imaging (RMI) is an effective technique to evaluate human tissue, cells and bodily fluids, including blood serum for disease diagnosis. ChemImage Corporation, in collaboration with clinicians, has been engaged in development of an in vitro diagnostic Raman assay focused on CRC detection. The Raman Assay for Colorectal Cancer (RACC) exploits the high specificity of Raman imaging to distinguish diseased from normal dried blood serum droplets without additional reagents. Pilot Study results from testing of hundreds of biobank patient samples have demonstrated that RACC detects CRC with high sensitivity and specificity. However, expanded clinical trials, which are ongoing, are revealing a host of important preanalytical considerations associated with sample collection, sample storage and stability, sample shipping, sample preparation and sample interferents, which impact detection performance. Results from recent clinical studies will be presented.
结直肠癌(CRC)是美国男性和女性中第三大常见癌症。拉曼分子成像(Raman Molecular Imaging, RMI)是一种评估人体组织、细胞和体液(包括血清)用于疾病诊断的有效技术。ChemImage公司与临床医生合作,致力于开发一种专注于CRC检测的体外诊断拉曼测定法。结直肠癌拉曼试验(RACC)利用拉曼成像的高特异性来区分病变和正常的干血清滴,而无需额外的试剂。对数百名生物库患者样本进行测试的初步研究结果表明,RACC检测结直肠癌具有高灵敏度和特异性。然而,正在进行的扩大临床试验揭示了与样品收集、样品储存和稳定性、样品运输、样品制备和样品干扰有关的许多重要的分析前考虑因素,这些因素会影响检测性能。将介绍最近临床研究的结果。
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引用次数: 0
Field-testing of a cost-effective mobile-phone based microscope for screening of Schistosoma haematobium infection (Conference Presentation) 一种具有成本效益的移动电话显微镜用于血球血吸虫感染筛查的现场测试(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2209697
Hatice Ceylan Koydemir, I. Bogoch, Derek K. Tseng, R. D. Ephraim, Evans Duah, Joseph Tee, J. Andrews, A. Ozcan
Schistosomiasis is a parasitic and neglected tropical disease, and affects <200-million people across the world, with school-aged children disproportionately affected. Here we present field-testing results of a handheld and cost effective smartphone-based microscope in rural Ghana, Africa, for point-of-care diagnosis of S. haematobium infection. In this mobile-phone microscope, a custom-designed 3D printed opto-mechanical attachment (~150g) is placed in contact with the smartphone camera-lens, creating an imaging-system with a half-pitch resolution of ~0.87µm. This unit includes an external lens (also taken from a mobile-phone camera), a sample tray, a z-stage to adjust the focus, two light-emitting-diodes (LEDs) and two diffusers for uniform illumination of the sample. In our field-testing, 60 urine samples, collected from children, were used, where the prevalence of the infection was 72.9%. After concentration of the sample with centrifugation, the sediment was placed on a glass-slide and S. haematobium eggs were first identified/quantified using conventional benchtop microscopy by an expert diagnostician, and then a second expert, blinded to these results, determined the presence/absence of eggs using our mobile-phone microscope. Compared to conventional microscopy, our mobile-phone microscope had a diagnostic sensitivity of 72.1%, specificity of 100%, positive-predictive-value of 100%, and a negative-predictive-value of 57.1%. Furthermore, our mobile-phone platform demonstrated a sensitivity of 65.7% and 100% for low-intensity infections (≤50 eggs/10 mL urine) and high-intensity infections (<50 eggs/10 mL urine), respectively. We believe that this cost-effective and field-portable mobile-phone microscope may play an important role in the diagnosis of schistosomiasis and various other global health challenges.
血吸虫病是一种被忽视的寄生性热带疾病,在全世界影响着不到2亿人,其中学龄儿童受到的影响尤为严重。在此,我们介绍了在非洲加纳农村使用的一种具有成本效益的手持式智能手机显微镜的现场测试结果,用于血链球菌感染的即时诊断。在这个手机显微镜中,一个定制设计的3D打印光学机械附件(~150g)与智能手机相机镜头接触,创建了一个半间距分辨率为~0.87µm的成像系统。该装置包括一个外部镜头(也来自移动电话相机),一个样品托盘,一个用于调整焦距的z级平台,两个发光二极管(led)和两个用于均匀照明样品的漫射器。在我们的现场测试中,使用了从儿童中收集的60份尿液样本,其中感染流行率为72.9%。在离心浓缩样品后,将沉积物放置在玻璃载玻片上,由专家诊断专家首先使用常规台式显微镜鉴定/定量haematobium虫卵,然后由另一位不知道这些结果的专家使用我们的手机显微镜确定虫卵的存在/不存在。与传统显微镜相比,我们的手机显微镜的诊断灵敏度为72.1%,特异性为100%,阳性预测值为100%,阴性预测值为57.1%。此外,我们的手机平台对低强度感染(≤50个鸡蛋/10毫升尿)和高强度感染(<50个鸡蛋/10毫升尿)的敏感性分别为65.7%和100%。我们相信,这种具有成本效益的便携式移动电话显微镜可能在血吸虫病和各种其他全球健康挑战的诊断中发挥重要作用。
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引用次数: 1
Exploring the nanoscale dynamics of biomolecules with optical microcavities (Conference Presentation) 利用光学微腔探索生物分子的纳米尺度动力学(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2220294
F. Vollmer
Medicine as well as biology increasingly rely on the use of cutting‐edge physics and engineering, in order to pursue the next generation nanomedical applications and to address fundamental questions in the life sciences. Central to this task is the study of micro- and nano systems, focusing on how engineered systems combined with natural ones can advance sensing, medicine, and our understanding of how biological systems work. My research addresses these important questions with state‐of‐the‐art biosensor technologies, capable of detecting single biomolecules and their dynamics; and resolving the kinetics of biomolecular systems on timescales ranging from few nanoseconds to several hours
医学和生物学越来越依赖于前沿物理学和工程学的应用,以追求下一代纳米医学的应用并解决生命科学中的基本问题。这项任务的核心是对微纳米系统的研究,重点关注工程系统如何与自然系统相结合,从而促进传感、医学和我们对生物系统如何工作的理解。我的研究用最先进的生物传感器技术解决了这些重要问题,能够检测单个生物分子及其动力学;并在从几纳秒到几个小时的时间尺度上解决生物分子系统的动力学问题
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引用次数: 0
Portable point-of-care blood analysis system for global health (Conference Presentation) 用于全球卫生的便携式医疗点血液分析系统(会议介绍)
Pub Date : 2016-05-03 DOI: 10.1117/12.2212666
J. Dou, J. S. Aitchison, Lu Chen, R. Nayyar
In this paper we present a portable blood analysis system based on a disposable cartridge and hand-held reader. The platform can perform all the sample preparation, detection and waste collection required to complete a clinical test. In order to demonstrate the utility of this approach a CD4 T cell enumeration was carried out. A handheld, point-of-care CD4 T cell system was developed based on this system. In particular we will describe a pneumatic, active pumping method to control the on-chip fluidic actuation. Reagents for the CD4 T cell counting assay were dried on a reagent plug to eliminate the need for cold chain storage when used in the field. A micromixer based on the active fluidic actuation was designed to complete sample staining with fluorescent dyes that was dried on the reagent plugs. A novel image detection and analysis algorithm was developed to detect and track the flight of target particles and cells during each analysis. The handheld, point-of-care CD4 testing system was benchmarked against clinical cytometer. The experimental results demonstrated experimental results were closely matched with the flow cytometry. The same platform can be further expanded into a bead-array detection system where other types of biomolecules such as proteins can be detected using the same detection system.
本文介绍了一种基于一次性血液盒和手持读取器的便携式血液分析系统。该平台可以完成临床试验所需的所有样品制备、检测和废物收集。为了证明这种方法的效用CD4 T细胞枚举进行。在此基础上开发了一种手持式、即时护理的CD4 T细胞系统。特别地,我们将描述一种气动、主动泵送方法来控制片上流体驱动。用于CD4 T细胞计数测定的试剂在试剂塞上干燥,以消除在现场使用时冷链储存的需要。设计了一种基于主动流体驱动的微混合器,用于用在试剂塞上干燥的荧光染料完成样品染色。提出了一种新的图像检测和分析算法,用于检测和跟踪每次分析过程中目标粒子和细胞的飞行。手持式,点护理CD4检测系统以临床细胞仪为基准。实验结果表明,实验结果与流式细胞术结果吻合较好。同样的平台可以进一步扩展成一个头阵列检测系统,其中其他类型的生物分子,如蛋白质,可以使用相同的检测系统进行检测。
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引用次数: 2
Fiber-optic Fourier transform infrared (FO-FTIR) spectroscopy for detecting endotoxin contamination in ophthalmic viscosurgical devices (OVDS) (Conference Presentation) 光纤傅里叶变换红外(FO-FTIR)光谱检测眼科粘手术器械(OVDS)内毒素污染(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2219746
Moinuddin Hassan, I. Ilev
Ophthalmic Viscosurgical Devices (OVDs) in clinical setting are a major health risk factor for potential endotoxin contamination in the eye, due to their extensive applications in cataract surgery for space creation, stabilization and protection of intraocular tissue and intraocular lens (IOL) during implantation. Endotoxin contamination of OVDs is implicated in toxic anterior syndrome (TASS), a severe complication of cataract surgery that leads to intraocular damage and even blindness. Current standard methods for endotoxin contamination detection utilize rabbit assay or Limulus amoebocyte lysate (LAL) assays. These endotoxin detection strategies are extremely difficult for gel-like type devices such as OVDs. To overcome the endotoxin detection limitations in OVDs, we have developed an alternative optical detection methodology for label-free and real-time sensing of bacterial endotoxin in OVDs, based on fiber-optic Fourier transform infrared (FO-FTIR) transmission spectrometry in the mid-IR spectral range from 2.5 micron to 12 micron. Endotoxin contaminated OVD test samples were prepared by serial dilutions of endotoxins on OVDs. The major results of this study revealed two salient spectral peak shifts (in the regions 2925 to 2890 cm^-1 and 1125 to 1100 cm^-1), which are associated with endotoxin in OVDs. In addition, FO-FTIR experimental results processed using a multivariate analysis confirmed the observed specific peak shifts associated with endotoxin contamination in OVDs. Thus, employing the FO-FTIR sensing methodology integrated with a multivariate analysis could potentially be used as an alternative endotoxin detection technique in OVD.
眼科粘手术装置(ovd)广泛应用于白内障手术中,用于在植入过程中创造空间、稳定和保护眼内组织和人工晶状体(IOL),是临床上造成眼内潜在内毒素污染的主要健康危险因素。ovd的内毒素污染与前毒性综合征(TASS)有关,前毒性综合征是白内障手术的一种严重并发症,可导致眼内损伤甚至失明。目前检测内毒素污染的标准方法采用兔法或鲎试剂(LAL)法。这些内毒素检测策略对于像ovd这样的凝胶型设备是非常困难的。为了克服ovd中细菌内毒素检测的局限性,我们开发了一种替代的光学检测方法,用于ovd中细菌内毒素的无标签实时检测,该方法基于光纤傅里叶变换红外(FO-FTIR)透射光谱,光谱范围为2.5微米至12微米。采用连续稀释的方法制备内毒素污染的OVD检测样品。本研究的主要结果揭示了两个显著的光谱峰移(在2925至2890 cm^-1和1125至1100 cm^-1区域),这与ovd的内毒素有关。此外,使用多变量分析处理的FO-FTIR实验结果证实了观察到的与内毒素污染相关的特定峰移。因此,将FO-FTIR传感方法与多变量分析相结合,有可能作为OVD的一种替代内毒素检测技术。
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引用次数: 0
Dual Raman-Brillouin microscope for chemical and mechanical characterization and imaging (Conference Presentation) 用于化学和机械表征和成像的双拉曼-布里渊显微镜(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2213691
V. Yakovlev
We present a unique confocal microscope capable of measuring the Raman and Brillouin spectra simultaneously from a single spatial location. Raman and Brillouin scattering offer complementary information about a material's chemical and mechanical structure, respectively, and concurrent monitoring of both of these spectra would set a new standard for material characterization. We achieve this by applying recent innovations in Brillouin spectroscopy that reduce the necessary acquisition times to durations comparable to conventional Raman spectroscopy while attaining a high level of spectral accuracy. To demonstrate the potential of the system, we apply this system for imaging melanoma, arteriosclerotic plaque, embryonic development, bones and biomaterials. The developed instrument has the potential for very diverse analytical applications in basic biomedical science and biomedical diagnostics and imaging.
我们提出了一个独特的共聚焦显微镜能够测量拉曼和布里渊光谱同时从一个单一的空间位置。拉曼散射和布里渊散射分别提供了关于材料化学和机械结构的补充信息,同时监测这两种光谱将为材料表征设定新的标准。我们通过应用布里渊光谱的最新创新来实现这一目标,将必要的采集时间减少到与传统拉曼光谱相当的持续时间,同时获得高水平的光谱精度。为了证明该系统的潜力,我们将该系统应用于黑色素瘤、动脉硬化斑块、胚胎发育、骨骼和生物材料的成像。所开发的仪器在基础生物医学科学和生物医学诊断和成像方面具有非常多样化的分析应用潜力。
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引用次数: 0
Engineering brighter probes for advanced cellular imaging (Conference Presentation) 用于高级细胞成像的工程更亮探针(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2225372
L. Lavis
The abstract is not available
摘要没有
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引用次数: 0
Phase-sensitive multiple reference optical coherence tomography (Conference Presentation) 相敏多参考光学相干层析成像(会议报告)
Pub Date : 2016-05-03 DOI: 10.1117/12.2213574
R. Dsouza, H. Subhash, K. Neuhaus, J. Hogan, Carol Wilson, M. Leahy
Multiple reference OCT (MR-OCT) is a recently developed novel time-domain OCT platform based on a miniature reference arm optical delay, which utilizes a single miniature actuator and a partial mirror to generate recirculating optical delay for extended axial-scan range. MR-OCT technology promises to fit into a robust and cost-effective design, compatible with integration into consumer-level devices for addressing wide applications in mobile healthcare and biometry applications. Using conventional intensity based OCT processing techniques, the high-resolution structural imaging capability of MR-OCT has been recently demonstrated for various applications including in vivo human samples. In this study, we demonstrate the feasibility of implementing phase based processing with MR-OCT for various functional applications such as Doppler imaging and sensing of blood vessels, and for tissue vibrography applications. The MR-OCT system operates at 1310nm with a spatial resolution of ~26 µm and an axial scan rate of 600Hz. Initial studies show a displacement-sensitivity of ~20 nm to ~120 nm for the first 1 to 9 orders of reflections, respectively with a mirror as test-sample. The corresponding minimum resolvable velocity for these orders are ~2.3 µm/sec and ~15 µm/sec respectively. Data from a chick chorioallantoic membrane (CAM) model will be shown to demonstrate the feasibility of MR-OCT for imaging in-vivo blood flow.
多参考OCT (MR-OCT)是近年来发展起来的一种基于微型参考臂光延迟的新型时域OCT平台,该平台利用单个微型驱动器和部分反射镜产生循环光延迟,以扩大轴向扫描范围。MR-OCT技术有望适应强大且经济高效的设计,兼容集成到消费者级设备中,以解决移动医疗和生物识别应用中的广泛应用。使用传统的基于强度的OCT处理技术,MR-OCT的高分辨率结构成像能力最近已被证明用于各种应用,包括体内人体样本。在这项研究中,我们证明了将基于相位的处理与MR-OCT应用于各种功能应用的可行性,如多普勒成像和血管传感,以及组织振动学应用。MR-OCT系统工作波长为1310nm,空间分辨率为~26µm,轴向扫描速率为600Hz。初步研究表明,以镜面为试样,前1 ~ 9阶反射的位移灵敏度分别为~20 ~ ~120 nm。这两阶对应的最小可分辨速度分别为~2.3µm/sec和~15µm/sec。来自小鸡绒毛尿囊膜(CAM)模型的数据将被证明MR-OCT用于体内血流成像的可行性。
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引用次数: 0
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SPIE BiOS
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