这项工作得到了国家眼科研究所 4K12EY022299 (YMP) 的慷慨支持, 为视力而战-国际视网膜研究基金会 FFS GIA16002 (YMP), 不受限制的部门支持从研究, 以防止失明, 和密歇根大学眼科和视觉科学系。这项工作利用了国家眼科研究所 P30 EY007003 资助的核心视觉研究中心。

兔子是美国农业部 (USDA) 覆盖的物种。它在生物医学研究中的应用需要遵循严格的规定。所有的兔实验都是按照帕特 (视觉和眼科研究协会) 在眼科和视力研究中使用动物的声明进行的, 这是在大学批准了实验动物协议后进行的。密歇根大学动物使用与护理委员会 (UCUCA) (协议 PRO00006486, PI Yannis)。
1. 系统配置
<ol><li>光声显微镜 (PAM)<ol>
<li>使用由二极管泵浦的固态激光器泵浦的光学参量振荡器 (参量) 作为 PAM 的光源. 选择合适的技术指标, 如脉冲重复率1赫、脉冲持续时间 3-6 ns 和可调谐波长范围 405-2600 nm。</li>
		<li>用两个镜 (M1 和 M2) 反射激光在 570 nm 的发射光束, 然后通过一个半波板衰减器安装在一个机动的旋转阶段, 最后聚焦, 过滤, 并准它由光束准直 (图 1)。优化光束准直器的设计。光束准直器的一个示例配置包括聚焦透镜 L1 (焦距 250 mm)、针孔 (直径50µm) 和准直透镜 L2 (焦距 30 mm)。</li>
		<li>将准直光束除以分光比为 90/10 (反射/透射) 的光束分配器 (BS1)。用光电二极管记录传输部分的脉冲-脉冲激光能量监测。连续偏转反射的部分由镜子 (M3) 和分色镜 (DM) 和光栅扫描它使用二维电流计。振镜是一个共享的组件与频谱域 (SD)-OCT 系统 (描述如下)。</li>
		<li>通过由扫描透镜 (焦距36毫米) 和眼科透镜 (OL, 焦距10毫米) 组成的望远镜传送扫描光束, 最后将其聚焦在兔眼光学的眼底。</li>
		<li>选择一个有适当技术规格的针形超声波换能器, 例如, 中心频率27兆赫, 双向 6 dB 带宽60%。将其与中央视轴上的结膜接触, 捕捉兴奋的光声信号。</li>
		<li>使用超声波放大器 (例如, 增益 57 dB) 放大信号, 通过低通滤波器 (例如, 截止频率 32 MHz) 对其进行过滤, 并以200毫秒/秒的采样速率将高速数字化器数字化。</li>
		<li>在兔眼上方放置一个电能表, 并测量在兔角膜上的激光脉冲能量, 使其低于 ANSI 安全限制160新泽西在 570 nm38。通过同步电子学, 用 Matlab 控制的激光快门来阻挡光束, 避免激光曝光。</li>
		<li>通过数据采集 (数据采集) 板同步激光器、振镜和数字化仪。编程软件进行系统控制和数据采集。</li>
	</ol>
</li>
	<li>光谱域光学相干层析成像 (SD OCT)<ol>
<li>在参考臂 (图 1) 中, 通过在扫描透镜 (SL) 和一块色散补偿玻璃 (DCG) 之后添加眼科透镜 (OL) 来适应基于商用系统的 SD OCT 系统。通过修改, OCT 系统可以实现对兔眼后段的图像处理。</li>
		<li>使用变焦外壳管调整参考臂的长度, 以确保它与样品臂的光学路径长度匹配。使用虹膜来控制反射光的亮度, 以确保其与背散射光强度的匹配, 从而达到最大的图像对比度。</li>
		<li>采用在扫描头上封装的电荷耦合器件 (CCD) 摄像机, 以照明发光二极管 (LED) 作为外部光源, 对兔眼底进行实时可视化。</li>
	</ol>
</li>
</ol>2. 系统对准
<ol><li>初始化电流计的位置, 并通过调整光纤准直器安装和分立方体的螺钉来对准 OCT 系统。 注: 分步程序可在商业获取的 OCT 系统手册中获得, 此处不包括。这一步骤主要是为了确保光纤准直器、参考臂和扫描透镜的正确对准, 以最大限度地提高 OCT 系统的性能。</li>
	<li>调整聚焦透镜焦点周围的针孔的x、 y和z位置, 以便在空间上滤除激光束, 并最大限度地传输激光能量。使用高度测量工具检查针孔前后的激光高度, 以确保它们是相同的。</li>
	<li>调整准直透镜 L2 的倾斜、偏心和z位置以准过滤光束。确保在近场和远场观测时, 光束的大小和高度大致相同。</li>
	<li>通过调整镜面和 DM 的倾斜, 将 PAM 激光光束和 OCT 光束进行共轴结合。在这一步之后, PAM 激光器和 OCT 光应该完全重合, 扫描区在兔眼底是相同的。</li>
	<li>调整 OL 透镜的倾斜和偏心, 使其在光学路径中正确对准。一旦完成, 双模态系统就可以成像了。 注: 你可以使用自动准直法来实现这一点, 即, 检查光线背面反射的 OL 透镜表面, 以确保它回去沿着同样的方式与入射光。</li>
</ol>3. 兔制剂
<ol><li>从动物设施中取一只新西兰白兔, 记录个人信息, 如体重和动物数量。</li>
	<li>山兔平台, 包括身体支持和头部支持的光学表下的成像系统。将水循环加热毯放在车身支撑上, 将循环水的温度设置为38° c, 以帮助保持家兔体温在实验和恢复期间的体温。</li>
	<li>记录手术前的生命体征, 包括总体动物状态、粘膜颜色、心率、呼吸速率和直肠体温。麻醉用氯胺酮 (40 毫克/千克) 和嗪 (5 毫克/千克) 的混合物通过肌肉注射和记录氯胺酮 (附表三控制物质) 的使用情况。通过检查心率、呼吸速率和整体状态来确认麻醉水平。</li>
	<li>放大兔学生使用一滴卡1% 眼科和肾上腺素盐酸2.5% 眼科。</li>
	<li>使用窥镜, 以保持眼睑的方式和应用一滴眼润滑剂滋润角膜。在成像程序之前, 在眼睛中灌输一滴局部丁卡因0.5%。 注: 对于动物可能不适的较长的程序或程序, 在实验前给兔子皮下注射昔以确保动物的舒适性。</li>
</ol>4. SD-OCT 成像
<ol><li>将兔子转移到临床眼底相机的成像平台上, 在 OCT 成像前进行50度眼底、无红色和自发荧光图像。这有助于检查眼睛的光学透明度和识别眼底血管的形态和标志, 如视神经和髓质视网膜血管。</li>
	<li>把兔子转移到 OCT 系统的平台上, 调整它的姿势, 大致定位在 OL 下的一只眼睛。用 LED 灯照亮眼睛。 注: 为方便临床翻译的技术, 兔眼不稳定使用任何其他方法和兔头是刚刚投入头部支持没有任何固定。</li>
	<li>打开 OCT 软件, 首先检查眼底的 CCD 摄像头图像。精细调整头部支持的高度和角度, 如果有必要, 以确保利益区 (roi), 如脉络膜血管和视网膜血管, 是在视野 (视) 的相机。 注意: 如果兔子处于良好的麻醉水平下, 一个连续的成像会话可以持续长达10分钟, 而无需重新调整兔头。</li>
	<li>画一条直线代表 OCT B 扫描兴趣并开始扫描。调整参考臂长度, 以可视化 oct 图像并优化 oct 软件中的色散补偿因子, 以获得最清晰的图像。 注意: 当调整参考臂长度时, 两个镜像的 OCT 图像将一个接一个地出现。正确的图像可以区分的基础上先验知识的眼底解剖。</li>
	<li>设置数据获取参数, 如像素数和平均值, 以及保存图像。</li>
	<li>观察家兔的呼吸速率和心率, 估计麻醉水平和动物舒适度。对于更长的疗程, 第三剂量的补充氯胺酮或吸入异氟醚可以考虑与气管插管, V 凝胶, 或口罩, 以维持在必要的麻醉平面。</li>
	<li>在实验中每2分钟用眼冲洗兔角膜, 以防止角膜表面脱水和角膜浅点上皮角膜。每15分钟监测和记录动物的生命体征。</li>
</ol>5. PAM 成像
<ol><li>戴上合适的激光安全护目镜, 打开参量激光器。</li>
	<li>启动 PAM 控制软件, 将激光波长调到目标团的一个吸收峰值 (例如, 570 nm 用于血红蛋白), 初始化电流计的位置, 并在兔角膜前监测激光能量, 以确保它低于 ANSI 安全限制。</li>
	<li>在三维 (3D) 平移阶段安装超声波换能器, 并将传感器尖端与指向眼底的兔结膜接触。使用一滴眼润滑剂更好地耦合传感器提示和兔结膜。</li>
	<li>打开 LED 照明灯, 通过 Matlab 软件对兔眼底进行可视化。</li>
	<li>设置扫描 ROI (即视网膜血管或脉络膜血管), 包括中心和物理尺寸。打开激光快门, 开始 B 扫描光束。通过大致校准传感器, 你应该能够看到在示波器上检测到的光声信号。如果没有, 稍微调整眼睛的位置, 以扫描不同区域的角膜或切换到另一只眼睛, 并重复上述过程。</li>
	<li>观察示波器上检测到的光声信号, 并微调传感器位置, 使整个 B 扫描信号强度最大化。 注: 由于光束宽度有限, 超声换能器通常有一个小视41。这一步决定了最终 PAM 图像的背景调制。不对称会导致 PAM 图像具有异构背景, 大大降低了图像质量。</li>
	<li>设置数据获取参数。这包括像素数 (e. g、256×256像素)、采样率 (e. g、200毫秒/秒) 和延迟时间。开始数据采集。当完成时, Matlab 软件将自动打开快门以通过激光束, 并关闭快门以阻止激光曝光。 注: 受激光脉冲重复率 (1 kHz) 的限制, 用 256 x 256 像素完成图像的数据采集需要大约1分钟。</li>
	<li>通过最大强度投影 (MIP)13或通过体积渲染38在3D 中处理原始数据并将 PAM 图像可视化为二维 (2D)。</li>
	<li>卸下超声波传感器, 用去离子水冲洗笔尖, 并将其放回存储箱。</li>
	<li>将兔子转移到眼底摄像机, 重新检查眼底。这一步有助于检查是否有任何形态学变化的眼底后, 成像会议。</li>
	<li>在实验中每两分钟用眼冲洗兔角膜, 以防止角膜表面脱水和角膜。每15分钟监测和记录动物的生命体征。 注: PAM、OCT 和眼底成像会议大约需要1小时。</li>
</ol>6. 后成像
<ol><li>眼底再检查后, 如果连接, 请断开 V 型凝胶。用眼冲洗眼睛, 应用芬眼和霉素, 素 B 硫酸盐和地塞米松眼膏, 闭上眼睛。</li>
	<li>将带水循环毯的兔子转移到回收室。把盒子从灯上屏蔽下来, 等到兔子自然苏醒。在此期间, 监测动物的生命体征每15分钟, 并保持记录, 并返回一份副本的动物设施, 保存。</li>
	<li>一旦兔子醒来, 是活跃的, 警觉和正常行走, 把它运回动物设施。如果一个急性实验计划, 安乐的动物使用安乐死解决方案 (e. g., Beuthanasia, 0.22 毫升/千克, 静脉注射在边缘耳静脉) 和处置的胴体。</li>
	<li>关闭软件和激光。清洁光学工作台。</li>
</ol>

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作者没有什么可透露的。

一个完整的和常规的泪膜是必不可少的高质量眼底图像。不规则和恶化的撕裂膜可以显著降低图像质量42。为了保持泪膜的完整性, 防止角膜表面有斑点的角膜, 重要的是用眼非常频繁地润滑角膜, 大约每两分钟。如果对眼睛的不透明度有任何顾虑, 请使用狭缝灯和荧光素条来检查角膜的状况。
较大的动物眼后段成像可能存在一些困难, 包括高频率分量、角膜?...

近几十年来见证了生物医学光声成像领域的爆炸性发展1,2,3,4,5,6,7 ,8。根据光的能量转换成声音, 新出现的微声成像可以可视化的生物样品从细胞器, 单元, 组织, 器官的小动物整个身体, 可以揭示其解剖, 功能, 分子, 遗传,和代谢信息1,2,9,10,11,12。光声成像在一系列生物医学领域中找到了独特的应用, 如细胞生物学13,14, 血管生物学15,16, 神经病学17,18, 肿瘤科19,20,21,22, 皮肤科23, 药理学24, 和血液病25,26。它在眼科的应用, 即光声眼部成像, 已经引起了科学家和临床医生的极大兴趣, 目前正在积极调查中。
与常规使用的眼部成像技术27不同, 如荧光血管造影 (FA) 和哚绿血管造影 (ICGA) (基于荧光对比), 光学相干层析 (OCT) (基于光学散射对比), 其衍生物 OCT 血管造影 (基于红细胞的运动对比), 光声眼成像以光学吸收为对比机制。这是不同于传统的眼科成像技术, 并提供了一个独特的工具, 研究光学吸收特性的眼睛, 这通常是与病理生理状态的眼部组织28。到目前为止, 在光声视觉成像中已经做了大量的出色工作29,30,31,323334 36,37, 但这些研究集中在小动物的眼球后段, 如老鼠和老鼠。开创性的研究充分证明了眼科光声成像的可行性, 但是, 由于大鼠和小鼠眼球尺寸小 (小于 1/3), 临床翻译技术还有很长的路要走。人类。由于超声波在较长距离上传播, 当使用这种技术来成像大眼睛后段时, 信号强度和图像质量会受到很大的影响。
为了达到这个目标, 我们最近报道了使用集成光声显微镜 (PAM) 和光谱域 oct (SD oct)38的活兔无创无标签脉络成像。该系统具有优异的性能, 可根据眼组织的内源吸收和散射对比, 对大动物眼的视网膜和脉络膜进行可视化。家兔的初步结果表明, PAM 可以非区分个别视网膜和脉络膜血管使用激光照射剂量 (〜80新泽西州) 显著低于美国国家标准学会 (ANSI) 安全限制 (160 新泽西州) 在570nm39;OCT 可以清楚地分辨出不同的视网膜层、脉络膜和巩膜。这是第一次演示的大动物后段成像, 使用 PAM, 可能是一个重要的步骤, 以临床翻译的技术考虑到兔眼球大小 (18.1 mm)40是几乎80% 的轴向长度人 (23.9 毫米)。
在这项工作中, 我们提供了一个详细的描述的双模态成像系统和实验协议, 用于无创, 无标签的脉络成像在活兔和演示系统性能通过代表性的视网膜和脉络膜成像结果。

<table border="0" cellpadding="0" cellspacing="0" style="width:824px;" width="822">
	<colgroup>
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		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dual-modality imaging system</td>
			<td style="width:279px;"></td>
			<td style="width:157px;"></td>
			<td style="width:159px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">OPO laser</td>
			<td style="width:279px;">Ekspla (Vilnius, Lithuania)</td>
			<td style="width:157px;">NT-242</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Beam attenuator</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AHWP10M-600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Motorized rotation stage</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">PRM1/MZ8</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Motorized rotation stage controller</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">TDC001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Focusing lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC254-250-B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Pinhole</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">P50S</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Collimating lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC127-030-B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Photodiode</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">PDA36A&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Laser shutter</td>
			<td style="width:279px;">Vincent Associates Inc. (Toronto, Canada)</td>
			<td style="width:157px;">LS6S2T0</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Laser shutter driver</td>
			<td style="width:279px;">Vincent Associates Inc. (Toronto, Canada)</td>
			<td style="width:157px;">VCM-D1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dichroic mirror</td>
			<td style="width:279px;">Semrock, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">Di03-R785-t3-25&#215;36</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Scan lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">OCT-LK3-BB</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Ophthalmic lens</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">AC080-010-B-ML</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Ultrasonic transducer</td>
			<td style="width:279px;">Optosonic Inc. (Arcadia, CA, USA)</td>
			<td style="width:157px;">Custom</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Amplifier</td>
			<td style="width:279px;">L3 Narda-MITEQ (Hauppauge, NY, USA)</td>
			<td style="width:157px;">AU-1647</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Band-pass filter</td>
			<td style="width:279px;">Mini-Circuits (Brooklyn, NY, USA)</td>
			<td style="width:157px;">BLP-30+</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Digitizer</td>
			<td style="width:279px;">DynamicSignals LLC (Lockport, IL, USA)</td>
			<td style="width:157px;">PX1500-4&#160;</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Synchronization electronics</td>
			<td style="width:279px;">National Instruments Corporation (Austin, TX, USA)</td>
			<td style="width:157px;">USB-6353</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">OCT module</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">Ganymede-II-HR</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Dispersion compensation glass</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">LSM03DC</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Illumination LED light</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">MCWHF2&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Power meter</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td style="width:157px;">S121C&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Power meter interface</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td>PM100USB&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Height measurement tool&#160;</td>
			<td style="width:279px;">Thorlabs, Inc. (Newton, NJ, USA)</td>
			<td>BHM1</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Fundus camera</td>
			<td style="width:279px;">Topcon Corporation (Tokyo, Japan)&#160;</td>
			<td>TRC 50EX</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Matlab</td>
			<td style="width:279px;">MathWorks (Natick, MA, USA)</td>
			<td style="width:157px;">2017a</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Oscilloscope</td>
			<td style="width:279px;">Teledyne LeCroy (Chestnut Ridge, NY, USA)</td>
			<td style="width:157px;">WaveJet 354T</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:229px;">Animal experiment</td>
			<td style="width:279px;"></td>
			<td style="width:157px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Water-circulating blanket</td>
			<td style="width:279px;">Stryker Corporation (Kalamazoo, MI, USA)</td>
			<td style="width:157px;">TP-700</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Ketamine hydrochloride injection</td>
			<td style="width:279px;">Par pharmaceutical, Inc. (Woodcliff Lake, NJ, USA)</td>
			<td style="width:157px;">NDC code 42023-115-10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Xylazine hydrochloride</td>
			<td>VetOne (Boise, ID, USA)</td>
			<td style="width:157px;">NDC code 13985-704-10</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Tropicamide ophthalmic</td>
			<td style="width:279px;">Akorn Pharmaceuticals Inc. (Lake Forest, IL, USA)</td>
			<td style="width:157px;">NDC code 17478-102-12</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Phenylephrine hydrochloride ophthalmic</td>
			<td style="width:279px;">Paragon BioTeck, Inc. (Portland, OR, USA)</td>
			<td style="width:157px;">NDC code 42702-102-15</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Eye lubricant</td>
			<td style="width:279px;">Hub Pharmaceuticals LLC (Rancho Cucamonga, CA, USA)</td>
			<td style="width:157px;">NDC code 17238-610-15</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Eyewash</td>
			<td style="width:279px;">Altaire Pharmaceuticals, Inc. (Aquebogue, NY, USA)</td>
			<td style="width:157px;">NDC code 59390-175-18</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Tetracaine hydrochloride ophthalmic solution</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-920-64</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Flurbiprofen sodium ophthalmic solution</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-314-25</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:229px;">Neomycin and Polymyxin B Sulfates and Dexamethasone Ophthalmic Ointment</td>
			<td style="width:279px;">Bausch &#38; Lomb, Inc. (Rochester, NY, USA)</td>
			<td style="width:157px;">NDC code 24208-795-35</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:229px;">Meloxicam injection</td>
			<td style="width:279px;">Henry Schein Inc. (Queens, NY, USA)</td>
			<td style="width:157px;">NDC code 11695-6925-1</td>
			<td></td>
		</tr>
	</tbody>
</table>

novel photoacoustic microscopy and optical coherence tomography dual-modality chorioretinal imaging in living rabbit eyes

光声眼部成像是一种新兴的眼科成像技术, 它可以通过将光能转换成声波来非可视化的眼部组织, 目前正在进行密集的调查。然而, 迄今为止报告的大部分工作都是针对小动物 (如大鼠和小鼠) 眼后段的成像, 这对由于眼球尺寸小而对临床人的翻译造成了挑战。这篇手稿描述了一个新的光声显微镜 (PAM) 和光学相干断层扫描 (OCT) 双模系统的后段成像的大动物的眼睛, 如兔。该系统配置, 系统对准, 动物准备, 和双模态实验协议为在体内, 无创, 无标签的脉络成像在兔子是详细的。该方法的有效性通过代表性的实验结果证明, 包括由 PAM 和 OCT 获得的视网膜和脉络膜血管。这篇手稿提供了一个实用的指南, 以再现兔的成像结果和推进大动物的光声眼部成像。

双模态成像系统和实验协议已成功地在作者的实验室使用四新西兰白兔进行了测试。下面展示了一些代表性的结果。
图 1显示了 PAM 和 SD OCT 双模态成像系统的示意图。它由以下几个模块组成: 光声光源、可变激光衰减器、光束准直仪、能量计、扫描头、光电探测和采集模块、OCT 单元和同步电子学。详细?...

Watch this Scientific Journal Video about Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes at JoVE.com

Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes

这份手稿描述了新的设置和操作程序的光声显微镜和光学相干断层扫描双模系统的无创, 无标签的脉络成像大动物, 如兔。

新型光声显微镜与光学相干层析成像在活兔眼中的双模脉络成像

Photoacoustic ocular imaging is an emerging ophthalmic imaging technology that can noninvasively visualize ocular tissue by converting light energy into ...

novel-photoacoustic-microscopy-optical-coherence-tomography-dual

Research

JoVE Journal

Engineering

10.8K 次观看.  University of Michigan. 这份手稿描述了新的设置和操作程序的光声显微镜和光学相干断层扫描双模系统的无创, 无标签的脉络成像大动物, 如兔。

视频: Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes

Construction and Testing of Coin Cells of Lithium Ion Batteries

Rechargeable lithium ion batteries have wide applications in electronics, where customers always demand more capacity and longer lifetime. Lithium ion batteries have also been considered to be used in electric and hybrid vehicles1 or even electrical grid stabilization systems2. All these applications simulate a dramatic increase in the research and development of battery materials3-7, including new materials3,8, doping9, nanostructuring10-13, coatings or surface modifications14-17 and novel binders18. Consequently, an increasing number of physicists, chemists and materials scientists have recently ventured into this area. Coin cells are widely used in research laboratories to test new battery materials; even for the research and development that target large-scale and high-power applications, small coin cells are often used to test the capacities and rate capabilities of new materials in the initial stage. 
 In 2010, we started a National Science Foundation (NSF) sponsored research project to investigate the surface adsorption and disordering in battery materials (grant no. DMR-1006515). In the initial stage of this project, we have struggled to learn the techniques of assembling and testing coin cells, which cannot be achieved without numerous help of other researchers in other universities (through frequent calls, email exchanges and two site visits). Thus, we feel that it is beneficial to document, by both text and video, a protocol of assembling and testing a coin cell, which will help other new researchers in this field. This effort represents the "Broader Impact" activities of our NSF project, and it will also help to educate and inspire students.
In this video article, we document a protocol to assemble a CR2032 coin cell with a LiCoO2 working electrode, a Li counter electrode, and (the mostly commonly used) polyvinylidene fluoride (PVDF) binder. To ensure new learners to readily repeat the protocol, we keep the protocol as specific and explicit as we can. However, it is important to note that in specific research and development work, many parameters adopted here can be varied. First, one can make coin cells of different sizes and test the working electrode against a counter electrode other than Li. Second, the amounts of C black and binder added into the working electrodes are often varied to suit the particular purpose of research; for example, large amounts of C black or even inert powder were added to the working electrode to test the "intrinsic" performance of cathode materials14. Third, better binders (other than PVDF) have also developed and used18. Finally, other types of electrolytes (instead of LiPF6) can also be used; in fact, certain high-voltage electrode materials will require the uses of special electrolytes7.

A protocol to construct and test coin cells of lithium ion batteries is described. The specific procedures of making a working electrode, preparing a counter electrode, assembling a cell inside a glovebox and testing the cell are presented.

钮扣电池锂离子电池的建设和测试

Rechargeable lithium ion batteries have wide applications in electronics, where customers always demand more capacity and longer lifetime.  Lithium ion ...

科研

工程学

Fabricating Metamaterials Using the Fiber Drawing Method

Metamaterials are man-made composite materials, fabricated by assembling components much smaller than the wavelength at which they operate 1. They owe their electromagnetic properties to the structure of their constituents, instead of the atoms that compose them. For example, sub-wavelength metal wires can be arranged to possess an effective electric permittivity that is either positive or negative at a given frequency, in contrast to the metals themselves 2. This unprecedented control over the behaviour of light can potentially lead to a number of novel devices, such as invisibility cloaks 3, negative refractive index materials 4, and lenses that resolve objects below the diffraction limit 5. However, metamaterials operating at optical, mid-infrared and terahertz frequencies are conventionally made using nano- and micro-fabrication techniques that are expensive and produce samples that are at most a few centimetres in size 6-7. Here we present a fabrication method to produce hundreds of meters of metal wire metamaterials in fiber form, which exhibit a terahertz plasmonic response 8. We combine the stack-and-draw technique used to produce microstructured polymer optical fiber 9 with the Taylor-wire process 10, using indium wires inside polymethylmethacrylate (PMMA) tubes. PMMA is chosen because it is an easy to handle, drawable dielectric with suitable optical properties in the terahertz region; indium because it has a melting temperature of 156.6 &deg;C which is appropriate for codrawing with PMMA. We include an indium wire of 1 mm diameter and 99.99% purity in a PMMA tube with 1 mm inner diameter (ID) and 12 mm outside diameter (OD) which is sealed at one end. The tube is evacuated and drawn down to an outer diameter of 1.2 mm. The resulting fiber is then cut into smaller pieces, and stacked into a larger PMMA tube. This stack is sealed at one end and fed into a furnace while being rapidly drawn, reducing the diameter of the structure by a factor of 10, and increasing the length by a factor of 100. Such fibers possess features on the micro- and nano- scale, are inherently flexible, mass-producible, and can be woven to exhibit electromagnetic properties that are not found in nature. They represent a promising platform for a number of novel devices from terahertz to optical frequencies, such as invisible fibers, woven negative refractive index cloths, and super-resolving lenses.

Metamaterials at terahertz frequencies offer unique opportunities, but are challenging to fabricate in bulk. We adapt the fabrication procedure for microstructured polymer optical fibers to inexpensively fabricate metamaterials potentially on an industrial scale. We produce polymethylmethacrylate fibers containing ~10 &mu;m diameter indium wires separated by ~100 &mu;m, which exhibit a terahertz plasmonic response.

制造超材料使用光纤拉丝法

Metamaterials are man-made composite materials, fabricated by assembling components much smaller than the wavelength at which they operate 1. They owe ...

Measurement of Coherence Decay in GaMnAs Using Femtosecond Four-wave Mixing

The application of femtosecond four-wave mixing to the study of fundamental properties of diluted magnetic semiconductors ((s,p)-d hybridization, spin-flip scattering) is described, using experiments on GaMnAs as a prototype III-Mn-V system.&#160; Spectrally-resolved and time-resolved experimental configurations are described, including the use of zero-background autocorrelation techniques for pulse optimization.&#160; The etching process used to prepare GaMnAs samples for four-wave mixing experiments is also highlighted.&#160; The high temporal resolution of this technique, afforded by the use of short (20 fsec) optical pulses, permits the rapid spin-flip scattering process in this system to be studied directly in the time domain, providing new insight into the strong exchange coupling responsible for carrier-mediated ferromagnetism.&#160; We also show that spectral resolution of the four-wave mixing signal allows one to extract clear signatures of (s,p)-d hybridization in this system, unlike linear spectroscopy techniques.&#160;&#160; This increased sensitivity is due to the nonlinearity of the technique, which suppresses defect-related contributions to the optical response. This method may be used to measure the time scale for coherence decay (tied to the fastest scattering processes) in a wide variety of semiconductor systems of interest for next generation electronics and optoelectronics.

The technique of femtosecond four-wave mixing is described, including spectrally-resolved and time-resolved configurations. We illustrate the utility of this technique for the investigation of crucial physical properties in the III-V diluted magnetic semiconductors, afforded by its nonlinearity and high temporal resolution.

The application of femtosecond four-wave mixing to the study of fundamental properties of diluted magnetic semiconductors ((s,p)-d hybridization, ...

Coherent anti-Stokes Raman Scattering (CARS) Microscopy Visualizes Pharmaceutical Tablets During Dissolution

Traditional pharmaceutical dissolution tests determine the amount of drug dissolved over time by measuring drug content in the dissolution medium. This method provides little direct information about what is happening on the surface of the dissolving tablet. As the tablet surface composition and structure can change during dissolution, it is essential to monitor it during dissolution testing. In this work coherent anti-Stokes Raman scattering microscopy is used to image the surface of tablets during dissolution while UV absorption spectroscopy is simultaneously providing inline analysis of dissolved drug concentration for tablets containing a 50% mixture of theophylline anhydrate and ethyl cellulose. The measurements showed that in situ CARS microscopy is capable of imaging selectively theophylline in the presence of ethyl cellulose. Additionally, the theophylline anhydrate converted to theophylline monohydrate during dissolution, with needle-shaped crystals growing on the tablet surface during dissolution. The conversion of theophylline anhydrate to monohydrate, combined with reduced exposure of the drug to the flowing dissolution medium resulted in decreased dissolution rates. Our results show that in situ CARS microscopy combined with inline UV absorption spectroscopy is capable of monitoring pharmaceutical tablet dissolution and correlating surface changes with changes in dissolution rate.

Coherent anti-Stokes Raman Scattering CARS Microscopy Visualizes Pharmaceutical Tablets During Dissolution

Coherent anti-Stokes Raman scattering (CARS) microscopy is combined with an intrinsic flow-through dissolution setup to allow in situ and real-time visualization of the surface of pharmaceutical tablets undergoing dissolution. Using this custom-built setup, it is possible to correlate CARS videos with drug dissolution profiles recorded using inline UV absorption spectroscopy.

相干反斯托克斯拉曼散射（CARS）显微可视化药片剂溶出过程中

Traditional pharmaceutical dissolution tests determine the amount of drug dissolved over time by measuring drug content in the dissolution medium. This ...

Writing Bragg Gratings in Multicore Fibers

Fiber Bragg gratings in multicore fibers can be used as compact and robust filters in astronomical and other research and commercial applications. Strong suppression at a single wavelength requires that all cores have matching transmission profiles. These gratings cannot be inscribed using the same method as for single-core fibers because the curved surface of the cladding acts as a lens, focusing the incoming UV laser beam and causing variations in exposure between cores. Therefore we use an additional optical element to ensure that the beam shape does not change while passing through the cross-section of the multicore fiber. This consists of a glass capillary tube which has been polished flat on one side, which is then placed over the section of the fiber to be inscribed. The laser beam enters the fiber through the flat surface of the capillary tube and hence maintains its original dimensions. This paper demonstrates the improvements in core-to-core uniformity for a 7-core fiber using this method. The technique can be generalized to larger multicore fibers.

We describe a technique for inscribing identical fiber Bragg gratings into each core of a multicore fiber. This is achieved by introducing an additional surface into the optical path to mitigate lensing by the curved surface of the fiber cladding.

写布拉格光栅在多核纤维

Fiber Bragg gratings in multicore fibers can be used as compact and robust filters in astronomical and other research and commercial applications. Strong ...

Compact Lens-less Digital Holographic Microscope for MEMS Inspection and Characterization

A micro-electro-mechanical-system (MEMS) is a widely used component in many industries, including energy, biotechnology, medical, communications, and automotive. However, effective inspection and characterization metrology systems are needed to ensure the functional reliability of MEMS. This study presents a system based on digital holography as a tool for MEMS metrology. Digital holography has gained increasing attention in the past 20 years. With the fast development and decreasing cost of sensor arrays, resolution of such systems has increased broadening potential applications. Thus, it has attracted attention from both research and industry sides as a potential reliable tool for industrial metrology. Indeed, by recording the interference pattern between an object beam (which contains sample height information) and a reference beam on a CCD camera, one can retrieve the quantitative phase information of an object. However, most of digital holographic systems are bulky and thus not easy to implement on industry production lines. The novelty of the system presented is that it is lens-less and thus very compact. In this study, it is shown that the Compact Digital Holographic Microscope (CDHM) can be used to evaluate several characteristics typically consider as criteria in MEMS inspections. The surface profiles of MEMS in both static and dynamic conditions are presented. Comparison with AFM is investigated to validate the accuracy of the CDHM.

We present a compact reflection digital holographic system (CDHM) for inspection and characterization of MEMS devices. A lens-less design using a diverging input wave providing natural geometrical magnification is demonstrated. Both static and dynamic studies are presented.

紧凑的镜头少数字全息显微镜的MEMS检测与表征

A micro-electro-mechanical-system (MEMS) is a widely used component in many industries, including energy, biotechnology, medical, communications, and ...

Dynamic Pore-scale Reservoir-condition Imaging of Reaction in Carbonates Using Synchrotron Fast Tomography

Underground storage permanence is a major concern for carbon capture and storage. Pumping CO2 into carbonate reservoirs has the potential to dissolve geologic seals and allow CO2 to escape. However, the dissolution processes at reservoir conditions are poorly understood. Thus, time-resolved experiments are needed to observe and predict the nature and rate of dissolution at the pore scale. Synchrotron fast tomography is a method of taking high-resolution time-resolved images of complex pore structures much more quickly than traditional &#181;-CT. The Diamond Lightsource Pink Beam was used to dynamically image dissolution of limestone in the presence of CO2-saturated brine at reservoir conditions. 100 scans were taken at a 6.1 &#181;m resolution over a period of 2 hours. The images were segmented and the porosity and permeability were measured using image analysis and network extraction. Porosity increased uniformly along the length of the sample; however, the rate of increase of both porosity and permeability slowed at later times.

Synchrotron fast tomography was used to dynamically image dissolution of limestone in the presence of CO2-saturated brine at reservoir conditions. 100 scans were taken at a 6.1 &#181;m resolution over a period of 2 h.

用同步快速反应层析成像动态孔隙规模油藏条件下成像碳酸盐

Underground storage permanence is a major concern for carbon capture and storage. Pumping CO2 into carbonate reservoirs has the potential to dissolve ...

In Depth Analyses of LEDs by a Combination of X-ray Computed Tomography (CT) and Light Microscopy (LM) Correlated with Scanning Electron Microscopy (SEM)

In failure analysis, device characterization and reverse engineering of light emitting diodes (LEDs), and similar electronic components of micro-characterization, plays an important role. Commonly, different techniques like X-ray computed tomography (CT), light microscopy (LM) and scanning electron microscopy (SEM) are used separately. Similarly, the results have to be treated for each technique independently. Here a comprehensive study is shown which demonstrates the potentials leveraged by linking CT, LM and SEM. In depth characterization is performed on a white emitting LED, which can be operated throughout all characterization steps. Major advantages are: planned preparation of defined cross sections, correlation of optical properties to structural and compositional information, as well as reliable identification of different functional regions. This results from the breadth of information available from identical regions of interest (ROIs): polarization contrast, bright and dark-field LM images, as well as optical images of the LED cross section in operation. This is supplemented by SEM imaging techniques and micro-analysis using energy dispersive X-ray spectroscopy.

In Depth Analyses of LEDs by a Combination of X-ray Computed Tomography CT and Light Microscopy LM Correlated with Scanning Electron Microscopy SEM

A workflow for comprehensive micro-characterization of active optical devices is outlined. It contains structural as well as functional investigations by means of CT, LM and SEM. The method is demonstrated for a white LED which can be still be operated during characterization.

在通过X射线计算机断层扫描（CT）和与扫描电子显微镜相关光学显微镜（LM）的一个组合的LED深度分析（SEM）的

In failure analysis, device characterization and reverse engineering of light emitting diodes (LEDs), and similar electronic components of ...

Optical Trap Loading of Dielectric Microparticles In Air

We demonstrate a method to trap a selected dielectric microparticle in air using radiation pressure from a single-beam gradient optical trap. Randomly scattered dielectric microparticles adhered to a glass substrate are momentarily detached using ultrasonic vibrations generated by a piezoelectric transducer (PZT). Then, the optical beam focused on a selected particle lifts it up to the optical trap while the vibrationally excited microparticles fall back to the substrate. A particle may be trapped at the nominal focus of the trapping beam or at a position above the focus (referred to here as the levitation position) where gravity provides the restoring force. After the measurement, the trapped particle can be placed at a desired position on the substrate in a controlled manner. 
In this protocol, an experimental procedure for selective optical trap loading in air is outlined. First, the experimental setup is briefly introduced. Second, the design and fabrication of a PZT holder and a sample enclosure are illustrated in detail. The optical trap loading of a selected microparticle is then demonstrated with step-by-step instructions including sample preparation, launching into the trap, and use of electrostatic force to excite particle motion in the trap and measure charge. Finally, we present recorded particle trajectories of Brownian and ballistic motions of a trapped microparticle in air. These trajectories can be used to measure stiffness or to verify optical alignment through time domain and frequency domain analysis. Selective trap loading enables optical tweezers to track a particle and its changes over repeated trap loadings in a reversible manner, thereby enabling studies of particle-surface interaction.

A protocol for launching and stably trapping selected dielectric microparticles in air is presented.

介电微粒的光阱装载在空气中

We demonstrate a method to trap a selected dielectric microparticle in air using radiation pressure from a single-beam gradient optical trap. Randomly ...

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy (ATOM)

Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the main trend in the advanced development of flow cytometry. Notably, adding high-resolution imaging capabilities allows for the complex morphological analysis of cellular/sub-cellular structures. This is not possible with standard flow cytometers. However, it is valuable for advancing our knowledge of cellular functions and can benefit life science research, clinical diagnostics, and environmental monitoring. Incorporating imaging capabilities into flow cytometry compromises the assay throughput, primarily due to the limitations on speed and sensitivity in the camera technologies. To overcome this speed or throughput challenge facing imaging flow cytometry while preserving the image quality, asymmetric-detection time-stretch optical microscopy (ATOM) has been demonstrated to enable high-contrast, single-cell imaging with sub-cellular resolution, at an imaging throughput as high as 100,000 cells/s. Based on the imaging concept of conventional time-stretch imaging, which relies on all-optical image encoding and retrieval through the use of ultrafast broadband laser pulses, ATOM further advances imaging performance by enhancing the image contrast of unlabeled/unstained cells. This is achieved by accessing the phase-gradient information of the cells, which is spectrally encoded into single-shot broadband pulses. Hence, ATOM is particularly advantageous in high-throughput measurements of single-cell morphology and texture &#8211; information indicative of cell types, states, and even functions. Ultimately, this could become a powerful imaging flow cytometry platform for the biophysical phenotyping of cells, complementing the current state-of-the-art biochemical-marker-based cellular assay. This work describes a protocol to establish the key modules of an ATOM system (from optical frontend to data processing and visualization backend), as well as the workflow of imaging flow cytometry based on ATOM, using human cells and micro-algae as the examples.

Microfluidic Imaging Flow Cytometry by Asymmetric-detection Time-stretch Optical Microscopy ATOM

This protocol describes the implementation of an asymmetric-detection time-stretch optical microscopy system for single-cell imaging in ultrafast microfluidic flow and its applications in imaging flow cytometry.

微流控成像流式细胞仪通过不对称检测时间延伸光学显微镜（ATOM）

Scaling the number of measurable parameters, which allows for multidimensional data analysis and thus higher-confidence statistical results, has been the ...