Name: multimodal volumetric retinal imaging by oblique scanning laser ophthalmoscopy (oslo) and optical coherence tomography (oct)
Uploaded: 2018-08-04T14:30:00
Description: Watch this Scientific Journal Video about Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT at JoVE.com

资金来源来自波士顿医疗中心的埃文斯医疗基金会提供的资金, 以及来自 NIH 5R01CA183101、CTSI 试点赠款1UL1TR001430、乔斯林试点项目和 CTSI KL2TR001411 的分包合同。

这里描述的所有方法都得到了波士顿医疗中心动物护理和使用委员会 (ACUC) 的批准。
1. 系统设置
<ol><li>奥斯陆系统<ol>
<li>使用超连续谱激光源作为系统激光源。<ol>
<li>用分色镜 (DM1) 将可见光范围 (450-650 nm) 从较高波长范围 (650-2000 nm) 中分离出来。在光束通过偏振光束分配器 (PBS) 后, 用一对色散棱镜展开频谱。</li>
			<li>放置一个狭缝, 以选择激发波长范围 (475-495 nm)。使用反射...

<ol>
	<li>Webb, R. H., Hughes, G. W., Delori, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Confocal+scanning+laser+ophthalmoscope.">Confocal scanning laser ophthalmoscope.</a> Applied Optics. 26 (8), 1492-1499 (1987).</li><li>Roorda, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adaptive+optics+scanning+laser+ophthalmoscopy.">Adaptive optics scanning laser ophthalmoscopy.</a> Optics Express. 10 (9), 405-412 (2002).</li><li>Huang, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography.">Optical coherence tomography.</a> Science. 254 (5035), 1178-1181 (1991).</li><li>de Carlo, T. E., Romano, A., Waheed, N. K., Duker, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+optical+coherence+tomography+angiography+(OCTA).">A review of optical coherence tomography angiography (OCTA).</a> International Journal of Retina and Vitreous. 1 (1), 5 (2015).</li><li>Jia, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+Optical+Coherence+Tomography+Angiography+of+Choroidal+Neovascularization+in+Age-Related+Macular+Degeneration.">Quantitative Optical Coherence Tomography Angiography of Choroidal Neovascularization in Age-Related Macular Degeneration.</a> Ophthalmology. 121 (7), 1435-1444 (2014).</li><li>Chen, C. -. L., Wang, R. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+coherence+tomography+based+angiography+[Invited].">Optical coherence tomography based angiography [Invited].</a> Biomedical Optics Express. 8 (2), 1056-1082 (2017).</li><li>Yi, J., Chen, S. Y., Shu, X., Fawzi, A. A., Zhang, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+retinal+imaging+using+visible-light+optical+coherence+tomography+guided+by+scanning+laser+ophthalmoscopy.">Human retinal imaging using visible-light optical coherence tomography guided by scanning laser ophthalmoscopy.</a> Biomedical Optics Express. 6 (10), 3701-3713 (2015).</li><li>Zhang, X. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual-band+spectral-domain+optical+coherence+tomography+for+in+vivo+imaging+the+spectral+contrasts+of+the+retinal+nerve+fiber+layer.">Dual-band spectral-domain optical coherence tomography for in vivo imaging the spectral contrasts of the retinal nerve fiber layer.</a> Optics Express. 19 (20), 19653-19667 (2011).</li><li>Delori, F. C., Webb, R. H., Sliney, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maximum+permissible+exposures+for+ocular+safety+(ANSI+2000),+with+emphasis+on+ophthalmic+devices.">Maximum permissible exposures for ocular safety (ANSI 2000), with emphasis on ophthalmic devices.</a> Journal of the Optical Society of America a-Optics Image Science and Vision. 24 (5), 1250-1265 (2007).</li><li>Zhang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volumetric+fluorescence+retinal+imaging+in+vivo+over+a+30-degree+field+of+view+by+oblique+scanning+laser+ophthalmoscopy+(oSLO).">Volumetric fluorescence retinal imaging in vivo over a 30-degree field of view by oblique scanning laser ophthalmoscopy (oSLO).</a> Biomedical Optics Express. 9 (1), 25-40 (2018).</li></ol>

在这里, 我们已经描述了奥斯陆, 一个在体内容积荧光视网膜成像技术与 FOV 超过了30°。与目前眼科护理成像方法标准的 OCT 相比, 奥斯陆提供了类似的3D 成像能力, 但也允许了 oct 不敏感的荧光对比度。奥斯陆的优势在于它只需要一个光栅扫描, 从而允许 OCT 的无缝组合, 为结构和荧光容积成像提供两种互补技术。
在该协议中, 获得良好图像质量的关键是大鼠眼的清晰度。...

眼科和视觉科学从现代光学成像技术中受益很大, 因为视网膜可以很容易地接触到光。荧光视网膜成像是诊断和治疗脉络膜视网膜血管疾病的重要工具, 如糖尿病视网膜病变 (DR) 和年龄相关的黄斑变性 (AMD), 这两者都是导致美国失明的主要原因。
然而, 利用荧光成像技术获得大视场 (FOV)、三维 (3D) 视网膜成像仍具有挑战性。眼底摄影没有深度分辨能力, 也不排斥漫射光。因此, 不同深度的信号混合可以降低图像质量。扫描激光检眼镜 (斯洛伐克) 和共焦斯洛伐克 (cSLO) 可通过共聚焦浇口1降低散射光的影响。然而, 由于其聚焦深度的限制, 斯洛伐克或 cSLO 很难获得3D 人的视网膜图像。自适应光学斯洛伐克 (AOSLO) 可以通过校正人眼所引入的波前畸变来提供高超的分辨率和对比度。然而, AOSLO 仍然需要 z 堆叠的容积成像2。光学相干层析成像 (oct)3和 oct 血管造影 (奥克塔) 系统克服这些限制, 提供三维 (3D) 解剖和血管图像 4,5,6, 但无染料的性质OCT 不能可视化泄漏指示血管功能障碍。
该协议描述了一种新的3D 容量荧光视网膜成像多模态平台, 即斜扫描激光检眼镜 (奥斯陆)。在这个成像系统中, 斜扫描是由鸽子尾滑块产生的, 最后的成像系统以角度对齐, 以检测荧光剖面图像。该系统采用激光扫描方法, 这些技术允许与 OCT 简单地结合, 作为一个互补的体积结构成像方式。目前, 大鼠视网膜的深度分辨率约为25µm, 视野为30°。从根本上说, 奥斯陆允许荧光版的 oct, 并可以同时结合 oct 和奥克塔在一个大 FOV。
在本议定书中, 我们将介绍奥斯陆的设置、对准和构造方法、大鼠视网膜活体成像方法和代表性结果。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Supercontinuum Laser Source</td>
			<td>NKT Photonics</td>
			<td>SuperK EXTREME EXU-OCT6</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichroic Mirror (DM1)</td>
			<td>Thorlabs</td>
			<td>DMLP650R</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichroic Mirror (DM2)</td>
			<td>Chroma</td>
			<td>ZT514/1064rpc</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichroic Mirror (DM3)</td>
			<td>Thorlabs</td>
			<td>DMLP900R</td>
			<td></td>
		</tr>
		<tr>
			<td>Single Mode Fiber (SMF 1)</td>
			<td>Thorlabs</td>
			<td>P3-460B-FC-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Single Mode Fiber (SMF 2)</td>
			<td>Thorlabs</td>
			<td>P3-780A-FC-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Optic Fiber Coupler</td>
			<td>Thorlabs</td>
			<td>TW850R5A2</td>
			<td></td>
		</tr>
		<tr>
			<td>1:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-100-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>3:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-150-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>3:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-50-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>1:1 Telescope System</td>
			<td>Thorlabs</td>
			<td>AC254-100-A&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>Galvo Mirrors (GM1,GM2)</td>
			<td>Thorlabs</td>
			<td>GVS201&#215;2</td>
			<td></td>
		</tr>
		<tr>
			<td>De-sacn Galvo Mirrors (GM3)</td>
			<td>Thorlabs</td>
			<td>GVS011</td>
			<td></td>
		</tr>
		<tr>
			<td>Objective Lens</td>
			<td>Olympus</td>
			<td>UplanSApo 20&#215;/0.75</td>
			<td></td>
		</tr>
		<tr>
			<td>Final imaging system</td>
			<td>Olympus</td>
			<td>UplanFL N 10&#215;/0.3</td>
			<td></td>
		</tr>
		<tr>
			<td>Final imaging system</td>
			<td>Computar</td>
			<td>12-36mm/1:2.8</td>
			<td></td>
		</tr>
		<tr>
			<td>Camera</td>
			<td>PCO</td>
			<td>Pco.pixelfly usb</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter</td>
			<td>Thorlabs</td>
			<td>FEL0800</td>
			<td></td>
		</tr>
		<tr>
			<td>Mounted Continuously Variable ND Filter</td>
			<td>Thorlabs</td>
			<td>NDC-50C-4M-A</td>
			<td></td>
		</tr>
		<tr>
			<td>Line Scan Camera</td>
			<td>Thorlabs</td>
			<td>SPL2048-140K</td>
			<td></td>
		</tr>
		<tr>
			<td>Analog Output Board (AO1)</td>
			<td>National Instrument</td>
			<td>PCI-6731</td>
			<td></td>
		</tr>
		<tr>
			<td>Analog Output Board (AO2)</td>
			<td>National Instrument</td>
			<td>PCIe-6351</td>
			<td></td>
		</tr>
		<tr>
			<td>Long pass filter</td>
			<td>Thorlabs</td>
			<td>FEL0800</td>
			<td></td>
		</tr>
	</tbody>
</table>

multimodal volumetric retinal imaging by oblique scanning laser ophthalmoscopy (oslo) and optical coherence tomography (oct)

虽然荧光成像在眼科广泛应用, 但大视野 (FOV) 三维 (3D) 荧光视网膜图像仍然是一个巨大的挑战与最新的视网膜成像方式, 因为他们需要 z 叠加编译容量数据集。新的光学相干层析成像 (oct) 和 oct 血管造影 (奥克塔) 系统克服了这些限制, 提供三维 (3D) 解剖和血管图像, 但 OCT 的无染料性质不能想象泄漏指示血管功能 障碍。该协议描述了一种新的斜扫描激光检眼镜 (奥斯陆) 技术, 提供3D 容量荧光视网膜成像。成像系统的设置产生斜扫描的鸽子尾巴滑块, 并使最终成像系统的角度, 以检测荧光横断面图像。该系统采用激光扫描方法, 因此, 允许简单地将 OCT 作为互补容积结构成像模式。对大鼠视网膜的活体成像进行了演示。荧光素溶液是静脉注射注射, 以产生容量荧光素血管造影 (vFA)。

图 4a显示了大鼠视网膜的横断面 OCT 图像。图 4b-4c显示同一时间获得的奥克塔和奥斯陆 vFA 相同的视网膜横断面图像。奥斯陆使横断面法类似于 OCT B 扫描。与奥克塔相比, 奥斯陆 vFA 横断面图像清楚地识别出神经纤维层 (NFL) 和神经节细胞层 (协鑫) 中的血管, 以及外层网状层 (OPL) 中的毛细血管。图 4d</s...

Watch this Scientific Journal Video about Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT at JoVE.com

Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy oSLO and Optical Coherence Tomography OCT

在这里, 我们提出了一个协议, 以获得一个大视野 (FOV) 三维 (3D) 荧光和 OCT 视网膜图像的新的成像多式联运平台。我们将介绍系统设置、校准方法和操作协议。在体内成像将被证明, 并有代表性的结果将提供。

斜扫描激光检眼镜 (奥斯陆) 与光学相干层析成像 (OCT) 的多模态视网膜成像

While fluorescence imaging is widely used in ophthalmology, a large field of view (FOV) three-dimensional (3D) fluorescence retinal image is still a big ...

Research

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Multimodal Volumetric Retinal Imaging by Oblique Scanning Laser Ophthalmoscopy (oSLO) and Optical Coherence Tomography (OCT)

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科研

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Both the clinical diagnosis and fundamental investigation of major ocular diseases greatly benefit from various non-invasive ophthalmic imaging ...

Wideband Optical Detector of Ultrasound for Medical Imaging Applications

宽带超声医疗成像应用，光学检测仪

Optical sensors of ultrasound are a promising alternative to piezoelectric techniques, as has been recently demonstrated in the field of optoacoustic ...

Biofunctionalized Prussian Blue Nanoparticles for Multimodal Molecular Imaging Applications

Biofunctionalized普鲁士蓝纳米粒子多式联运分子成像应用

Multimodal, molecular imaging allows the visualization of biological processes at cellular, subcellular, and molecular-level resolutions using multiple, ...

Computed Tomography and Optical Imaging of Osteogenesis-angiogenesis Coupling to Assess Integration of Cranial Bone Autografts and Allografts

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A major parameter determining the success of a bone-grafting procedure is vascularization of the area surrounding the graft. We hypothesized that ...

Nondestructive Monitoring of Degradable Scaffold-Based Tissue-Engineered Blood Vessel Development Using Optical Coherence Tomography

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Engineered vascular grafts with structural and mechanical properties similar to natural blood vessels are expected to meet the growing demand for arterial ...

Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells

Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy Conpokal on Live Cells

近同步激光扫描在活细胞上的共和和原子力显微镜（康波卡尔）

Techniques available for micro- and nano-scale mechanical characterization have exploded in the last few decades. From further development of the scanning ...

Dual Raster-Scanning Photoacoustic Small-Animal Imager for Vascular Visualization

用于血管可视化的双栅格扫描光声小动物成像仪

Imaging of vascular networks on small animals has played an important role in basic biomedical research. Photoacoustic imaging technology has great ...

In vivo Structural Assessments of Ocular Disease in Rodent Models using Optical Coherence Tomography

使用光学相干断层扫描对啮齿动物模型中眼部疾病的体内结构评估

Spectral-domain optical coherence tomography (SD-OCT) is useful for visualizing retinal and ocular structures in vivo. In research, SD-OCT is a valuable ...

Quantifying Fibrillar Collagen Organization with Curvelet Transform-Based Tools

使用基于曲线的转换工具量化纤维胶原蛋白组织

Fibrillar collagens are prominent extracellular matrix (ECM) components, and their topology changes have been shown to be associated with the progression ...