这项研究是由美国国立卫生研究院授予RO1 013516和赠款形式Optovue的支持。

1。协议文本<ol><li>患者被扫描RTVue傅立叶域光学相干断层成像（OCT）系统（公司Optovue，弗里蒙特，CA，美国）使用的circumpapillary的双圆形扫描（CDCS）和3D模型视盘扫描。 <ol><li> CDCS图案周围的视神经头由两个同心圆。内圈直径为3.40毫米，环的外直径为3.75毫米。这种模式横断所有的视网膜分支动脉和静脉产生的视神经头。双圈在单次扫描覆盖约2个心动周期的6倍。为了计算，多普勒频移和多普勒流速将估计对船只在OCT图像检测。 （ 图1a，图1b） </li><li> A“双角”的协议，用于获取多普勒OCT扫描。在“双角”协议中，3次扫描得到的OCT光束通过的瞳孔和3次扫描的的superonasal部通过inferonasal陈健波<html> TION。 </li><li>每次扫描的质量进行评估信号强度，运动误差和多普勒角度。技术人员需要重新做任何扫描，没有通过质量检查。每只眼睛进行6可接受的CDCS扫描，总共。 “双角”协议确保了每艘船舶，至少有一半的扫描提供了良好的多普勒角度。 </li><li> 3D碟片扫描模式是一个光栅扫描覆盖6x6毫米视盘周围地​​区。它仅进行一次，并提供了一​​个详细的连接这个区域的人脸图像。 （ 图1b） </li><li>也是进口的彩色照片视盘，以帮助区分动脉和静脉。 </li></ol></li></ol><img alt="图1" src="/files/ftp_upload/3524/3524fig1a.jpg" /> 图1a。的双圆circumpapillary扫描和3D磁盘扫描，并使用DOCTORC的的总血流量。 PHA“&gt; <li>多普勒角是该船只的探测光束与法线之间的角度，和多普勒信号的多普勒频移探测光束的轴线平行的流速分量成正比。因此流速可以估算，从多普勒角度和多普勒信号。然而，这两种多普勒角和多普勒信号变得不可靠的，当它们是低于噪声电平。另一方面，当多普勒角大时，多普勒频移将满分的可测量范围内。当多普勒角是合适的，在这项研究中使用的FD-OCT系统约5-15度，流速可以正确估计的多普勒频移和角度。 </li></ol><img alt="图1" src="/files/ftp_upload/3524/3524fig1b.jpg" /> 图1b。 <ol start="2" type="a"><li>在每个容器中的多普勒角测量由容器管腔的相对位置在t禾同心圆形扫描。的轴向分量的流速是通过以下方式获得的多普勒频移测定从相邻的的华侨城轴向扫描的相对相位。的总的流速来计算从轴向流分量和多普勒角。每个容器中的流动，然后通过集成超过血管横截面面积的总的速度更新计算。匹配连接面部图像的3D磁盘扫描和眼底照片，以确定每个容器无论是作为静脉或动脉的血管图案。视网膜总流量的计算方法是总结在视网膜静脉， 即 ，那些管腔直径33微米以上的流动。 </li></ol><ol start="2"><li>使用RTVue软件的原始数据导出为所有的扫描相同的眼睛。原始的OCT图像，包括多普勒和强度的图像，首先对图像质量测试使用“多普勒光学相干断层扫描视网膜循环”软件（DOCTORC）。 <ol><li>眼球运动测量ured内界膜和两帧之间的最大差值，估计由散装议案由合并的标准偏差。 </li><li>信号强度（反射率和估算的平均多普勒角）也计算。 </li><li>根据平均眼球运动和信号强度的重复扫描，对数据进行分类是“好”或“差”的质量方针。只有高质量的数据进行分级。 </li></ol></li><li>对于可接受的扫描，自动分割算法船舶检测施加到每个OCT图像。 <ol><li>匹配船只一个自动化的算法，该算法是用来定位在每个帧中的同一容器。注册在同一圆上的帧中，以及在内圈和外圈的两个平均帧创建。反射率和多普勒图像被平均。 </li><li>的概述，将检测到的容器投射到连接面从3D碟片扫描计算的眼底图像的线段。 </李&gt; </ol></li><li>平地机评论平均的内圈和外圈用一个圆圈表示自动分割结果重叠的帧的一小部分上的每个容器。 <ol><li>年级的法官的位置，血管直径，血管型（静脉/动脉）相匹配的船只上的两个环是正确的根据OCT形象的，恩面对OCT图像和光盘相同的眼睛的照片。 </li><li>平地机允许改变上述的任何值，如果他认为这是必要的。平地机也判断容器中的多普勒信号的质量，并给出了一个主观的置信度得分为每个扫描容器标签上。 </li><li>多普勒信号强度的血管面积的基础上，以每艘船舶被自动赋予一个置信度得分为0-5。然后手动校正年级的学生的实力血管多普勒信号，规律性的容器边界，容器大小的内圈和外圈之间的协议的基础上，签订协议的DOP在内圈和外圈之间的更简单移位。 </li></ol></li><li>后，所有船只进行验证和修正，自动算法计算每根血管的血流量，改编自我们以前发布的方法。 <ol><li>多普勒信号集成在血管区之间的所有帧，然后平均。然后，计算的求和的多普勒信号的多普勒角除以多普勒血流。 </li><li>对于每个容器，6的扫描评价主观置信度分数，多普勒角度，和变异系数的多普勒角度。如果超过1扫描通过质量检查，静脉的流动被认为是有效的。对于通过的质量检查的血管时，流程有效扫描之间的平均值。 </li><li>对于容器的无效结果，流是使用从有效的静脉血管面积和平均流量速度估计。平均流动速度的计算方法是在有效的静脉并除以流求和总结领域的人脉。流量的估计从平均流量速度开始，然后校正流速对血管面积的依赖。较大的血管有较高的平均流速。因此，根据修正后的斜率平均速度与血管直径2.13，即我们之前所报道。 </li><li>有效的静脉和无效静脉的估计流量的计算流量被添加，以确定总的视网膜血流。 </li><li>总的视网膜血流评估结果的基础上有效的静脉面积百分比，眼球运动，和信号强度。 </li><li>静脉面积和总动脉面积也得到加入容器区域。假设全视网膜血流量在动脉和静脉，动脉和静脉的速度计算，除以动脉的面积和静脉区的血流。 </li></ol></li></ol> 2。代表性的成果<p类=“jove_content”&gt;普通和青光眼的选择青光眼研究（AIGS， <a href="http://www.aigstudy.net" target="_blank">www.aigstudy.net</a> ）的先进的成像。 48眼进行扫描的“双角”协议和生产的扫描，通过图像质量检查。使用DOCTORC软件，有效的流量测量，可以得到83％的眼睛。 为了评估的重复性的DOCTORC系统，另一个较小的数据集有20只眼进行分级3年级的学生。这个数据集也被用来训练和测试年级的学生。 2年级学生使用的半自动化DOCTORC的软件和1使用在以前的出版物中的较早的完全手动的软件。2,3的总的视网膜血流量（ 表1）由两个年级学生使用DOCTORC软件确定是彼此相似的，并流入其他年级的学生使用手册软件的价格确定。只有65％的眼睛有有效的结果，因为有些的T他的数据没有根据双角协议，但单角协议。2单角协议包括5个多普勒扫描OCT光束通过瞳孔中心。因此，多普勒角度是更频繁地小，因此，较大部分的船只通常是不进行分级。 对于所有年级的学生，跨年级的学生可重复性，变异系数作为衡量的，是类似青光眼与正常眼（ 表2）。同样地，再现性测量的两种方法，DOCTORC和软件的操作手册，1-5是相似的（ 表2）。对于三年级学生，良好的相关性之间存在着总的血流量和从视场试验（ 图2），用于青光眼的图案的标准偏差。 <table border="1"><tbody><tr><td></td><td colspan="3"> DOCTORC软件手册软件3 </TD&gt; </tr><tr><td>条件</td><td>平地机1 </td><td>平地机2 </td><td></td></tr><tr><td>正常</td><td> 47.0±9.1 </td><td> 48.7±7.2 </td><td> 48.0±6.5 </td></tr><tr><td>青光眼</td><td> 36.5±5.5 </td><td> 36.7±5.9 </td><td> 34.9±5.1 </td></tr></tbody></table> 表1。视网膜全血流量使用2个不同的软件。 <table border="1"><tbody><tr><td></td><td colspan="2"> 变异系数 </td></tr><tr><td>青光眼（7只眼） </td><td></td><td></td></tr><tr><td>平地机1比平地机2（DOCTORC） </td><td> 9.58％ </td><td></td></tr><tr><td> DOCTORC与手动方法3 </td><td></td><td></td></tr><tr><td>平地机1 </td><td> 8.00％ </td><td></td> &lt;/ TR&gt; <tr><td>平地机2 </td><td> 9.74％ </td><td></td></tr><tr><td> 正常（6只眼） </td><td></td><td></td></tr><tr><td>平地机1比平地机2（DOCTORC） </td><td> 5.99％ </td><td></td></tr><tr><td> DOCTORC与手动方法</td><td></td><td></td></tr><tr><td>平地机1 </td><td> 8.87％ </td><td></td></tr><tr><td>平地机2 </td><td> 9.98％ </td><td></td></tr></tbody></table> 表2。总视网膜血流测量的可重复性。 <img alt="图2" src="/files/ftp_upload/3524/3524fig2.jpg" /> 图2，总视网膜血流和视野在青光眼的相关性。平地机1 DOCTORC软件。视野缺损总结模式标准差（P = 0.048），B平地机2使用DOCTORC软件。视野缺损总结，模式标准差（P = 0.032）。

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黄医生收到的资金支持，专利使用费，股票期权，交通费支援费，演讲费从Optovue，公司，谭智和王医生收取专利使用费和资金支持从Optovue，公司博士Koduru博士和Sadda收到的资金支持从Optovue。

血流量异常发生在青光眼的视网膜血管性疾病，如糖 ​​尿病性视网膜病变。6-10体积测量视网膜的血流量提供了有价值的信息4-6,11,12 DOCTORC提供了一个实用的方法来估计全视网膜疾病的进程。血流量的测量值的基础上使用的双圆圈扫描图案由多普勒OCT确定个别船只。1-5 平均总视网膜血流量测量多普勒OCT在正常的眼睛是47-49μl/ min的，与文献值34-65μl/ mi...

<table border="1"><tbody><tr><td> 的试剂/设备名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td> RTVue傅立叶域光学相干断层扫描</td><td> Optovue </td><td> N / A </td><td> 6.1.0.21或更高版本 安装与血流量的双环形扫描模式</td></tr></tbody></table>

doppler optical coherence tomography of retinal circulation

傅立叶域光学相干断层扫描（OCT）系统使用一个circumpapillary双循环扫描（CDCS），它可以扫描周围的视神经头3.40毫米和3.75毫米直径的非接触式视网膜血流进行测量。双同心圆连续进行了6倍，超过2秒。 CDCS扫描保存与多普勒频移信息可以计算出从该流。标准的临床协议要求3 CDCS扫描的OCT光束通过的superonasal瞳孔边缘3 CDCS扫描通过inferonal瞳孔。这双角协议，确保可接受的每个视网膜分支血管多普勒角度上获得至少1次扫描。 CDCS扫描数据，一个3 - 维体积OCT扫描的视盘扫描，和视盘的彩色照片一起使用，以获得上的眼睛的视网膜血流测量。我们已经开发出一种血流量测量软件被称为“多普勒光纤人相干断层扫描视网膜循环“（DOCTORC）。此半自动化的软件被用来测量视网膜全血流量，血管的横截面面积，和平均血流速度，每个容器中的流是从容器截面的多普勒频移计算截面面积和多普勒角之间的容器中，在OCT束。共有视网膜血流量测量从视盘周围静脉相加得到的结果在我们的多普勒OCT读取中心平地机和方法（&lt;10％之间表现出良好的再现性，OCT在青光眼患者视网膜血流测量高度与视野缺损（R 2&gt; 0.57，视野模式偏差），总视网膜血流可能是有用的管理，青光眼，视网膜疾病，视网膜疾病。）多普勒OCT是一种新的方法来进行快速，非接触式和可重复的测量使用广泛可傅立叶域OCT instrumentatio的的总视网膜血流的。N。这项新技术可以改善的临床研究和常规临床实践中进行这些测量的实用性。

Watch this Scientific Journal Video about Doppler Optical Coherence Tomography of Retinal Circulation at JoVE.com

Doppler Optical Coherence Tomography of Retinal Circulation

总的测量视网膜血流多普勒光学相干断层扫描和半自动化分级软件。

多普勒光学相干断层扫描视网膜循环

Noncontact retinal blood flow measurements are performed with a Fourier domain optical coherence tomography (OCT) system using a circumpapillary double ...

doppler-optical-coherence-tomography-of-retinal-circulation

Research

JoVE Journal

Medicine

18.6K 次观看.  Oregon Health and Science University. 总的测量视网膜血流多普勒光学相干断层扫描和半自动化分级软件。

视频: Doppler Optical Coherence Tomography of Retinal Circulation

Near Infrared Optical Projection Tomography for Assessments of &beta;-cell Mass Distribution in Diabetes Research

By adapting OPT to include the capability of imaging in the near infrared (NIR) spectrum, we here illustrate the possibility to image larger bodies of pancreatic tissue, such as the rat pancreas, and to increase the number of channels (cell types) that may be studied in a single specimen. We further describe the implementation of a number of computational tools that provide: 1/ accurate positioning of a specimen's (in our case the pancreas) centre of mass (COM) at the axis of rotation (AR)2; 2/ improved algorithms for post-alignment tuning which prevents geometric distortions during the tomographic reconstruction2 and 3/ a protocol for intensity equalization to increase signal to noise ratios in OPT-based BCM determinations3. In addition, we describe a sample holder that minimizes the risk for unintentional movements of the specimen during image acquisition. Together, these protocols enable assessments of BCM distribution and other features, to be performed throughout the volume of intact pancreata or other organs (e.g. in studies of islet transplantation), with a resolution down to the level of individual islets of Langerhans.

Near Infrared Optical Projection Tomography for Assessments of &amp;#946;-cell Mass Distribution in Diabetes Research

We describe the adaptation of optical projection tomography (OPT)1 to imaging in the near infrared spectrum, and the implementation of a number of computational tools. These protocols enable assessments of pancreatic &beta;-cell mass (BCM) in larger specimens, increase the multichannel capacity of the technique and increase the quality of OPT data.

近红外光学投影层析成像技术在糖尿病研究β细胞质量分布的评估

By adapting OPT to include the capability of imaging in the near infrared (NIR) spectrum, we here illustrate the possibility to image larger bodies of ...

科研

医学

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

生成和动脉病变的小鼠的三维定量使用光学投影层析成像

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Minimally Invasive Thumb-sized Pterional Craniotomy for Surgical Clip Ligation of Unruptured Anterior Circulation Aneurysms

Less invasive surgical approaches for intracranial aneurysm clipping may reduce length of hospital stay, surgical morbidity, treatment cost, and improve patient outcomes. We present our experience with a minimally invasive pterional approach for anterior circulation aneurysms performed in a major tertiary cerebrovascular center and compare the results with an aged matched dataset from the Nationwide Inpatient Sample (NIS). From August 2008 to December 2012, 22 elective aneurysm clippings on patients &#8804;55 years of age were performed by the same dual fellowship-trained cerebrovascular/endovascular neurosurgeon. One patient (4.5%) experienced transient post-operative complications. 18 of 22 patients returned for follow-up imaging and there were no recurrences through an average duration of 22 months. A search in the NIS database from 2008 to 2010, also for patients aged &#8804;55 years of age, yielded 1,341 hospitalizations for surgical clip ligation of unruptured cerebral aneurysms. Inpatient length of stay and hospital charges at our institution using the minimally invasive thumb-sized pterional technique were nearly half that of NIS (length of stay: 3.2 vs 5.7 days; hospital charges: $52,779 vs. $101,882). The minimally invasive thumb-sized pterional craniotomy allows good exposure of unruptured small and medium-sized supraclinoid anterior circulation aneurysms. Cerebrospinal fluid drainage from key subarachnoid cisterns and constant bimanual microsurgical techniques avoid the need for retractors which can cause contusions, localized venous infarctions, and post-operative cerebral edema at the retractor sites. Utilizing this set of techniques has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.

Minimally invasive thumb-sized pterional craniotomy for aneurysm clipping has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.

微创拇指大小翼点入开颅未破裂前循环动脉瘤的手术夹子结扎

Less invasive surgical approaches for intracranial aneurysm clipping may reduce length of hospital stay, surgical morbidity, treatment cost, and improve ...

In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses

Optical coherence tomography (OCT) is the optical equivalent of ultrasound imaging, based on the backscattering of near infrared light. OCT provides real time images with a 15 &#181;m axial resolution at an effective tissue penetration of 2-3 mm. Within the OCT images the loss of signal intensity per millimeter of tissue penetration, the attenuation coefficient, is calculated. The attenuation coefficient is a tissue specific property, providing a quantitative parameter for tissue differentiation. 
Until now, renal mass treatment decisions have been made primarily on the basis of MRI and CT imaging characteristics, age and comorbidity. However these parameters and diagnostic methods lack the finesse to truly detect the malignant potential of a renal mass. A successful core biopsy or fine needle aspiration provides objective tumor differentiation with both sensitivity and specificity in the range of 95-100%. However, a non-diagnostic rate of 10-20% overall, and even up to 30% in SRMs, is to be expected, delaying the diagnostic process due to the frequent necessity for additional biopsy procedures. 
We aim to develop OCT into an optical biopsy, providing real-time imaging combined with on-the-spot tumor differentiation. This publication provides a detailed step-by-step approach for percutaneous, needle based, OCT of renal masses.

In Vivo, Percutaneous, Needle Based, Optical Coherence Tomography of Renal Masses

Optical coherence tomography (OCT) is a high resolution imaging technique that allows analysis of tissue specific optical properties providing the means for tissue differentiation. We developed needle based OCT, providing real-time imaging combined with on-the-spot tumor differentiation. This publication describes a method for percutaneous, needle based OCT of renal masses.

在体内 ，经皮，基于针，肾群众的光学相干断层扫描

Optical coherence tomography (OCT) is the optical equivalent of ultrasound imaging, based on the backscattering of near infrared light. OCT provides real ...

Fluorescent Dye Labeling of Erythrocytes and Leukocytes for Studying the Flow Dynamics in Mouse Retinal Circulation

The retinal and choroidal blood flow dynamics may provide insight into the pathophysiology and sequelae of various ocular diseases, such as glaucoma, diabetic retinopathy, age-related macular degeneration (AMD) and other ocular inflammatory conditions. It may also help to monitor the therapeutic responses in the eye. The proper labeling of the blood cells, coupled with live-cell imaging of the labeled cells, allows for the investigation of the flow dynamics in the retinal and choroidal circulation. Here, we describe the standardized protocols of 1.5% indocyanine green (ICG) and 1% sodium fluorescein labeling of mice erythrocytes and leukocytes, respectively. Scanning laser ophthalmoscopy (SLO) was applied to visualize the labeled cells in the retinal circulation of C57BL/6J mice (wild type). Both methods demonstrated distinct fluorescently labeled cells in the mouse retinal circulation. These labeling methods can have wider applications in various ocular disease models.

Live-cell imaging of the labeled blood cells in ocular circulation can provide information about inflammation and ischemia in diabetic retinopathy and age-related macular degeneration. A protocol to label blood cells and image the labeled cells in the retinal circulation is described.

用于研究小鼠视网膜循环中流动动力学的红细胞和白细胞的荧光染料标记

The retinal and choroidal blood flow dynamics may provide insight into the pathophysiology and sequelae of various ocular diseases, such as glaucoma, ...

Ultrahigh Resolution Mouse Optical Coherence Tomography to Aid Intraocular Injection in Retinal Gene Therapy Research

HR-SD-OCT is utilized to monitor the progression of photoreceptor degeneration in live mouse models, assess the delivery of therapeutic agents into the subretinal space, and to evaluate toxicity and efficacy in vivo. HR-SD-OCT uses near infrared light (800-880 nm) and has optics specifically designed for the unique optics of the mouse eye with sub-2-micron axial resolution. Transgenic mouse models of outer retinal (photoreceptor) degeneration and controls were imaged to assess the disease progression. Pulled glass microneedles were used to deliver sub retinal injections of adeno-associated virus (AAV) or nanoparticles (NP) via a trans-scleral and trans-choroidal approach. Careful positioning of the needle into the subretinal space was required prior to a calibrated pressure injection, which delivers fluid into the sub retinal space. Real time subretinal surgery was conducted on our retinal imaging system (RIS). HR-SD-OCT demonstrated progressive uniform retinal degeneration due to expression of a toxic mutant human mutant rhodopsin (P347S) (RHOP347S) transgene in mice. HR-SD-OCT allows rigorous quantification of all the retinal layers. Outer nuclear layer (ONL) thickness and photoreceptor outer segment length (OSL) measurements correlate with photoreceptor vitality, degeneration, or rescue. The RIS delivery system allows real-time visualization of subretinal injections in neonatal (~P10-14) or adult mice, and HR-SD-OCT immediately determines success of delivery and maps areal extent. HR-SD-OCT is a powerful tool that can evaluate the success of subretinal surgery in mice, in addition to measuring vitality of photoreceptors in vivo. HR-SD-OCT can also be used to identify uniform animal cohorts to evaluate the extent of retinal degeneration, toxicity, and therapeutic rescue in preclinical gene therapy research studies.

Here we demonstrate a novel approach to using high resolution spectral-domain optical coherence tomography (HR-SD-OCT) to assist delivery of gene therapy agents into the subretinal space, assess its areal coverage, and characterize photoreceptor vitality.

超高分辨率小鼠光学相干层析成像辅助眼内注射在视网膜基因治疗中的应用

HR-SD-OCT is utilized to monitor the progression of photoreceptor degeneration in live mouse models, assess the delivery of therapeutic agents into the ...

In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy

Spectral domain optical coherence tomography (SD-OCT) and scanning laser ophthalmoscopy (SLO) are extensively used in experimental ophthalmology. In the present protocol, mice expressing green fluorescent protein (gfp) under the promoter of Cx3cr1 (BALB/c-Cx3cr1gfp/gfp) were used to image microglia cells in vivo in the retina. Microglia are resident macrophages of the retina and have been implicated in several retinal diseases1,2,3,4,5,6. This protocol provides a detailed approach for generation of retinal B-scans, with SD-OCT, and imaging of microglia cell distribution in Cx3cr1gfp/gfp mice with SLO in vivo, using an ophthalmic imaging platform system. The protocol can be used in several reporter mouse lines. However, there are some limitations to the protocol presented here. First, both SLO and SD-OCT, when used in the high-resolution mode, collect data with high axial resolution but the lateral resolution is lower (3.5 &#181;m and 6 &#181;m, respectively). Moreover, the focus and saturation level in SLO is highly dependent on parameter selection and correct alignment of the eye. Additionally, using devices designed for human patients in mice is challenging due to the higher total optical power of the mouse eye compared to the human eye; this can lead to lateral magnification inaccuracies7, which are also dependent on the magnification by the mouse lens among others. However, despite that the axial scan position is dependent upon lateral magnification, the axial SD-OCT measurements are accurate8.

In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy

This protocol describes how high-resolution imaging techniques such as spectral domain optical coherence tomography and scanning laser ophthalmoscopy can be utilized in small rodents, using an ophthalmic imaging platform system, to obtain information on retinal thickness and microglial cell distribution, respectively.

在体内图像的Cx3cr1gfp/gfp 记者光谱域光学相干层析成像和扫描激光眼底

Spectral domain optical coherence tomography (SD-OCT) and scanning laser ophthalmoscopy (SLO) are extensively used in experimental ophthalmology. In the ...

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

As a functional status of microcirculation, microvascular vasomotion is important for the delivery of oxygen and nutrients and the removal of carbon dioxide and waste products. The impairment of microvascular vasomotion might be a crucial step in the development of microcirculation-related diseases. In addition, the highly vascularized pancreatic islet is adapted to support endocrine function. In this respect, it seems possible to infer that the functional status of pancreatic islet microvascular vasomotion might affect pancreatic islet function. Analyzing the pathological changes of the functional status of pancreatic islet microvascular vasomotion may be a feasible strategy to determine contributions that pancreatic islet microcirculation makes to related diseases, such as diabetes mellitus, pancreatitis, etc. Therefore, this protocol describes using a laser Doppler blood flow monitor to determine the functional status of pancreatic islet microvascular vasomotion, and to establish parameters (including average blood perfusion, amplitude, frequency, and relative velocity of pancreatic islet microvascular vasomotion) for evaluation of the microcirculatory functional status. In a streptozotocin-induced diabetic mouse model, we observed an impaired functional status of pancreatic islet microvascular vasomotion. In conclusion, this approach for assessing pancreatic islet microvascular vasomotion in vivo may reveal mechanisms relating to pancreatic islet diseases.

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

Pancreatic islet microvascular vasomotion regulates islet blood distribution and maintains the physiological function of islet &#946; cells. This protocol describes using a laser Doppler monitor to determine the functional status of pancreatic islet microvascular vasomotion in vivo and to assess the contributions of pancreatic islet microcirculation to pancreatic-related diseases.

激光多普勒: 一种测量胰岛微血管血管舒缩体内的工具

As a functional status of microcirculation, microvascular vasomotion is important for the delivery of oxygen and nutrients and the removal of carbon ...

Evaluation of Cerebral Blood Flow Autoregulation in the Rat Using Laser Doppler Flowmetry

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained using laser Doppler flowmetry (LDF). This paper demonstrates a closed skull preparation that allows cerebral blood flow to be assessed without penetrating the skull or installing a chamber or cerebral window. To evaluate autoregulatory mechanisms, a model of controlled blood pressure reduction via graded hemorrhage can be utilized while simultaneously employing LDF. This enables the real time tracking of the relative changes in the blood flow in response to reductions in arterial blood pressure produced by the withdrawal of circulating blood volume. This paradigm is a valuable approach to study cerebral blood flow autoregulation during reductions in arterial blood pressure and, with minor modifications in the protocol, is also valuable as an experimental model of hemorrhagic shock. In addition to evaluating autoregulatory responses, LDF can be used to monitor the cortical blood flow when investigating metabolic, myogenic, endothelial, humoral, or neural mechanisms that regulate cerebral blood flow and the impact of various experimental interventions and pathological conditions on cerebral blood flow.

This article demonstrates the use of laser Doppler flowmetry to evaluate the ability of the cerebral circulation to autoregulate its blood flow during reductions in arterial blood pressure.

使用激光多普勒流测量测量大鼠脑血流自动调节的评估

When investigating the body&#39;s mechanisms for regulating cerebral blood flow, a relative measurement of microcirculatory blood flow can be obtained ...

Laser Doppler Perfusion Imaging in the Mouse Hindlimb

Blood flow recovery is a critical outcome measure after experimental hindlimb ischemia or ischemia-reperfusion. Laser Doppler perfusion imaging (LDPI) is a common, noninvasive, repeatable method for assessing blood flow recovery. The technique calculates overall blood flow in the sampled tissue from the Doppler shift in frequency caused when a laser hits moving red blood cells. Measurements are expressed in arbitrary perfusion units, so the contralateral non-intervened upon leg is usually used to help control measurements. Measurement depth is in the range of 0.3-1 mm; for hindlimb ischemia, this means that dermal perfusion is assessed. Dermal perfusion is dependent on several factors&#8212;most importantly skin temperature and anesthetic agent, which must be carefully controlled to result in reliable readings. Furthermore, hair and skin pigmentation can alter the ability of the laser to either reach or penetrate to the dermis. This article demonstrates the technique of LDPI in the mouse hindlimb.

Here, we present a protocol that demonstrates the technique and necessary controls for Laser Doppler perfusion imaging to measure blood flow in the mouse hindlimb.

激光多普勒灌注成像在鼠标欣德利姆

Blood flow recovery is a critical outcome measure after experimental hindlimb ischemia or ischemia-reperfusion. Laser Doppler perfusion imaging (LDPI) is ...