Name: switchable acoustic and optical resolution photoacoustic microscopy for in vivo small-animal blood vasculature imaging
Uploaded: 2017-06-26T15:00:00
Description: Watch this Scientific Journal Video about Switchable Acoustic and Optical Resolution Photoacoustic Microscopy for In Vivo Small-animal Blood Vasculature Imaging at JoVE.com

作者要感谢新加坡教育部（ARC2 / 15：M4020238）资助的二级拨款的财政支持。作者还要感谢周伟荣博比机车店的帮助。

所有动物实验均按照新加坡南洋理工大学机构动物保护和使用委员会（动物标本编号ARF-SBS / NIE-A0263）的批准规定和指导方针进行。 AR-OR-PAM系统（ 图1 ） <ol><li> 系统配置：AR-PAM <ol><li>使用由二极管泵浦的固态Nd-YAG激光器（532nm）和可调谐范围为559-576nm的染料激光器组成的纳秒可调谐激光器系统作为光学照射源。使用外部控制器将激光波长?...

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总而言之，已经开发出可切换的AR和OR PAM系统，其可以在更低的成像深度处实现高分辨率成像并且在较高的成像深度处实现较低分辨率的成像。确定可切换系统的横向分辨率和成像深度。该可切换PAM系统的优点包括：（1）使用紧密光学聚焦的高分辨率成像; （2）使用声学聚焦的深层组织成像; 3）AR-PAM的暗场照明，防止强烈的PA信号出现在皮肤表面; 4）将样品保持在一个地方，而不是在不同系统之?...

高分辨率光学成像方式，如光学相干断层扫描，共聚焦显微镜和多光子显微镜，有许多好处。然而，随着成像深度的增加，空间分辨率显着降低。这是因为软组织中光传输的漫射性质1,2 。光激发和超声检测的集成提供了一种解决方案，以克服深层组织中高分辨率光学成像的挑战。光声显微镜（PAM）是一种可以提供比其他光学成像模式更深的成像的方式。已成功应用于体内结构，功能，分子和细胞成像3,4,5,6,7,8 </sup&gt; ， 9，10，11，12，13研究结合强光学吸收对比度与超声波的高空间分辨率。 在PAM中，短的激光脉冲照射组织/样品。通过发色团（ 例如，黑色素，血红蛋白，水等 ）的光吸收导致温度升高，这又导致以声波（光声波）的形式产生压力波。产生的光声波可以由组织边界外的宽带超声换能器检测。利用弱光学和紧密的声学聚焦，可以在声分辨光声显微镜（AR-PAM） 14,15,16中实现深层组织成像。在AR-PAM，横向分辨率为45μm，成像深度高达3mm，已被证明15 。为了在声学上解析单个毛细管（约5μm），需要在&gt; 400 MHz中心频率下运行的超声波换能器。在这样高的频率下，穿透深度小于100μm。使用紧密的光学聚焦可以解决紧密的聚焦造成的问题。光学分辨率光声显微镜（OR-PAM）能够分辨单个毛细管，甚至单个细胞17 ，并且已经实现了0.5μm的横向分辨率18,19,20,21,22,23,24 。使用光子纳米喷嘴可以帮助实现超出衍射极限分辨率的分辨率n 25，26 。在OR-PAM中，由于聚焦而使穿透深度受到限制，并且它可以在生物组织23内形象高达〜1.2mm 。因此，AR-PAM可以使图像更深，但分辨率更低，OR-PAM可以以非常高的分辨率进行成像，但成像深度有限。 AR和OR-PAM系统的成像速度主要取决于激光源27的脉冲重复率。 结合AR-PAM和OR-PAM将对需要高分辨率和深度成像的应用非常有利。将这些系统结合在一起已经做了很少的努力。通常，使用两种不同的成像扫描仪进行成像，这要求样品在两个系统之间移动，因此难以进行体内成像。然而，使用AR和OR PAM的混合成像可实现具有可扩展分辨率a的成像深度。在一种方法中，使用光纤束来传送用于AR和OR PAM的光。在这种方法中，使用两个单独的激光器（对于AR为570nm的高能激光器，对于OR为532nm的低能量，高重复率激光器），使得系统不方便和昂贵28 。 OR-PAM激光波长是固定的，并且使用该组合系统不可能进行许多研究，例如氧饱和度。 AR和OR PAM之间的比较研究也是不可能的，因为AR和OR之间的激光波长不同。此外，AR-PAM使用明场照明;因此，来自皮肤表面的强光电信号限制了图像质量。因此，该系统不能用于许多生物成像应用。在另一种执行AR和OR PAM的方法中，光学和超声波聚焦被移动，这使得光焦点和超声波聚焦不对齐。因此，图像质量不是最佳的<sup class =“xref”&gt; 29。使用这种技术，AR-PAM和OR-PAM可以分别达到139μm和21μm的分辨率，使其分辨率不高。据报道另一种方法是改变光纤和准直光学器件，以便在AR和OR PAM之间进行切换，使得对准过程困难。在所有这些情况下，AR-PAM都不使用暗场照明。暗场照明的使用可以减少从皮肤表面产生强烈的光声信号。因此，使用环形照明可以进行深层组织成像，因为深色光声信号的检测灵敏度将比亮场照明的检测灵敏度高。 这项工作报告了一个可切换的AR和OR PAM（AR-OR-PAM）成像系统，能够对相同样品进行高分辨率成像和低分辨率深层组织成像，使用相同的激光和扫描仪进行两个系统EMS。 AR-OR-PAM系统的性能通过使用幻影实验确定空间分辨率和成像深度来表征。 在小鼠耳朵上进行体内血液血管成像以证明其生物成像能力。

<table border="0" cellpadding="0" cellspacing="0" width="939">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Q-switched Nd:YAG laser</td>
			<td style="width:232px;">Edgewave</td>
			<td style="width:177px;">BX80-2-L</td>
			<td style="width:227px;">Pump laser&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Credo-High Repetition Rate Dye Laser</td>
			<td>Spectra physics</td>
			<td style="width:177px;">CREDO-DYE-N</td>
			<td>Dye laser</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Precision Linear Stage</td>
			<td>Physik Instrumente</td>
			<td>PLS 85&#160;</td>
			<td>XY raster scanning stage</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Translation stage</td>
			<td>Physik Instrumente</td>
			<td>VT 80&#160;</td>
			<td>Confocal determine</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mounted Silicon photodiode</td>
			<td>Thorlabs</td>
			<td>SM05PD1A</td>
			<td>Triggering/Pulse variation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Motorized continuous Rotational stage&#160;</td>
			<td>Thorlabs</td>
			<td>CR1/M-Z7</td>
			<td>Diverting laser beam</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mounted Continuously Variable ND Filter</td>
			<td>Thorlabs</td>
			<td>NDC-50C-4M</td>
			<td>Intensity variable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fiber Patch Cable</td>
			<td>Thorlabs</td>
			<td>M29L01</td>
			<td>Multimode fiber</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Microscope objective</td>
			<td>Newport</td>
			<td>M-10X</td>
			<td>Objective&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">XY translating mount</td>
			<td>Thorlabs</td>
			<td>CXY1</td>
			<td>Translating mount</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Plano convex lens</td>
			<td>Thorlabs</td>
			<td>LA1951</td>
			<td>Collimating lens</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Conical lens&#160;</td>
			<td>Altechna</td>
			<td>APX-2-B254</td>
			<td>Ring shape beam</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Translation stage</td>
			<td>Thorlabs</td>
			<td>CT1</td>
			<td>Translating stage</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Optical condenser</td>
			<td style="width:232px;">Home made</td>
			<td style="width:177px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ultrasonic transducer</td>
			<td>Olympus-NDT</td>
			<td>V214-BB-RM</td>
			<td>50MHz transducer</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Plano concave lens</td>
			<td>Thorlabs</td>
			<td>LC4573</td>
			<td>Acoustic lens</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pulser/Receiver</td>
			<td>Olympus-NDT</td>
			<td>5073PR</td>
			<td>Pulse echo amplifier&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Mounted standard iris</td>
			<td>Thorlabs</td>
			<td>ID12/M</td>
			<td>Beam shaping</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Plano convex lens</td>
			<td>Thorlabs</td>
			<td>LA4327</td>
			<td>Condenser lens</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Mounted precision pinhole</td>
			<td>Thorlabs</td>
			<td>P50S</td>
			<td>Spatial filtering</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Single mode fiber patch cable</td>
			<td>Thorlabs</td>
			<td>P1-460B-FC-1</td>
			<td>Single mode fiber</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Fiber coupler</td>
			<td>Newport</td>
			<td>F-91-C1</td>
			<td>Single mode coupling</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Achromatic doublet lens</td>
			<td style="width:232px;">Edmund Optics</td>
			<td>32-317</td>
			<td>Achromatic doublet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Protected silver elliptical mirror</td>
			<td>Thorlabs</td>
			<td>PFE10-P01</td>
			<td>Mirror</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Right angle kinematic mirror mount</td>
			<td>Thorlabs</td>
			<td>KCB1</td>
			<td>Mirror mount</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Z-Axis Translation Mount</td>
			<td>Thorlabs</td>
			<td>SM1Z</td>
			<td>z translator</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Lens tube</td>
			<td>Thorlabs</td>
			<td>SM05L10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">UV Fused Silica Right-Angle Prism</td>
			<td>Thorlabs</td>
			<td>PS615</td>
			<td>Right angle prism</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Rhomboid prism</td>
			<td style="width:232px;">Edmund Optics</td>
			<td>47-214</td>
			<td>Shear wave</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dimethylpolysiloxane</td>
			<td>Sigma Aldrich</td>
			<td>DMPS1M</td>
			<td>Silicon oil</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Amplifier</td>
			<td>Mini Circuits</td>
			<td>ZFL-500LN</td>
			<td>Amplifier</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">16 bit high speed digitizer</td>
			<td>Spectrum</td>
			<td>M4i.4420</td>
			<td>Data acquisition card</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Oscilloscope</td>
			<td style="width:232px;">Agilent Technologies</td>
			<td style="width:177px;">DS06014A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Mice&#160;</td>
			<td style="width:232px;">InVivos Pte.Ltd</td>
			<td style="width:177px;">ICR</td>
			<td>Animal model</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Ultrasound gel&#160;</td>
			<td>Progress/parker acquasonic gel</td>
			<td>PA-GEL-CLEA-5000</td>
			<td>Acoustic coupling</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Water tank</td>
			<td style="width:232px;">Home made</td>
			<td style="width:177px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Translation stage</td>
			<td style="width:232px;">Homemade</td>
			<td style="width:177px;"></td>
			<td>Switching AR-OR</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Gold nanoparticles</td>
			<td>Sigma Aldrich</td>
			<td align="right">742031</td>
			<td>Lateral resolution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Sterile ocular ointment</td>
			<td style="width:232px;">Alcon</td>
			<td style="width:177px;">Duratears</td>
			<td>Animal imaging</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">1951 USAF resolution test target</td>
			<td style="width:232px;">Edmund Optics</td>
			<td align="right" style="width:177px;">38257</td>
			<td>Confocal alignment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Data acquisition software</td>
			<td style="width:232px;">National Instrument</td>
			<td style="width:177px;">Labview</td>
			<td>Home made software using Labview</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:303px;">Image Processing software</td>
			<td style="width:232px;">Mathworks</td>
			<td style="width:177px;">Matlab</td>
			<td>Home made program using Matlab</td>
		</tr>
	</tbody>
</table>

switchable acoustic and optical resolution photoacoustic microscopy for in vivo small-animal blood vasculature imaging

光声显微镜（PAM）是一种快速增长的invivo成像模式，结合光学和超声波，提供高分辨率穿透超过光学平均自由程（皮肤约1毫米）。通过将光学吸收对比度与单一模态中超声的高空间分辨率相结合，该技术可以穿透深层组织。光声显微镜系统可以具有较低的声学分辨率和探针深度或较高的光学分辨率和探针。通过单个系统实现高空间分辨率和大深度穿透是具有挑战性的。这项工作提出了一个AR-OR-PAM系统，能够在浅层深度进行高分辨率成像和体内相同样品的低分辨率深层组织成像。使用光学聚焦的1.4mm成像深度的横向分辨率为4μm，使用声学聚焦的具有7.8mm成像深度的45μm的横向分辨率成功利用组合系统进行演示。在这里， 进行体内小动物血液血管成像以证明其生物成像能力。

AR-OR-PAM系统的原理图如图1所示。在这种设置中，所有组件都集成在一起，并组装在一个光学保持架中。使用笼式系统可使AR-OR-PAM扫描头紧凑，易于组装，对齐和集成到单个扫描台上。 在图像采集期间使用成像头的二维连续光栅扫描。时间分辨的PA信号乘以声速（1,540m / s）以获得A线。在Y阶段的连续运动?...

Watch this Scientific Journal Video about Switchable Acoustic and Optical Resolution Photoacoustic Microscopy for In Vivo Small-animal Blood Vasculature Imaging at JoVE.com

Switchable Acoustic and Optical Resolution Photoacoustic Microscopy for In Vivo Small-animal Blood Vasculature Imaging

这里展示了可在体内在相同样品上进行浅层深度高分辨率成像和低分辨率深层组织成像的可切换声学分辨率（AR）和光学分辨率（OR）光声显微镜（AR-OR-PAM）系统。

可切换声光学分辨率光声显微镜<em&gt;体内</em&gt;小动物血液血管成像

Photoacoustic microscopy (PAM) is a fast-growing invivo imaging modality that combines both optics and ultrasound, providing penetration beyond the ...

Research

JoVE Journal

Bioengineering

Switchable Acoustic and Optical Resolution Photoacoustic Microscopy for In Vivo Small-animal Blood Vasculature Imaging

Three-dimensional Optical-resolution Photoacoustic Microscopy

三维光学分辨率的光声显微镜

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As ...

科研

生物工程

Integrated Photoacoustic Ophthalmoscopy and Spectral-domain Optical Coherence Tomography

集成光声眼底镜检查和谱域光学相干断层扫描

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 ...

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

受体动力学通过单分子荧光显微镜的高分辨率时空分析

Single-molecule microscopy is emerging as a powerful approach to analyze the behavior of signaling molecules, in particular concerning those aspect (e.g., ...

Preparation of Mica Supported Lipid Bilayers for High Resolution Optical Microscopy Imaging

支持的脂双层云母制备的高分辨率光学显微成像

Supported lipid bilayers (SLBs) are widely used as a model for studying membrane properties (phase separation, clustering, dynamics) and its interaction ...

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

Fluorescence-quenching of a Liposomal-encapsulated Near-infrared Fluorophore as a Tool for In Vivo Optical Imaging

荧光淬一个脂质体包封的近红外荧光团的作为一个工具<em&gt;在体内</em&gt;光学成像

Optical imaging offers a wide range of diagnostic modalities and has attracted a lot of interest as a tool for biomedical imaging. Despite the enormous ...

A High-performance Compact Photoacoustic Tomography System for In Vivo Small-animal Brain Imaging

A High-performance Compact Photoacoustic Tomography System for In Vivo Small-animal Brain Imaging

高性能紧凑型光声层析成像系统<em&gt;体内</em&gt;小动物脑成像

In vivo small-animal imaging has an important role to play in preclinical studies. Photoacoustic tomography (PAT) is an emerging hybrid imaging modality ...

Hand-held Clinical Photoacoustic Imaging System for Real-time Non-invasive Small Animal Imaging

手持式实时非侵入性小动物成像的临床光声成像系统

Translation of photoacoustic imaging into the clinic is a major challenge. Handheld real-time clinical photoacoustic imaging systems are very rare. Here, ...

Rigid Embedding of Fixed and Stained, Whole, Millimeter-Scale Specimens for Section-free 3D Histology by Micro-Computed Tomography

用微计算机断层扫描法对无节段3D 组织学进行固定染色、全尺寸、毫米度试样的刚性嵌入

For over a hundred years, the histological study of tissues has been the gold standard for medical diagnosis because histology allows all cell types in ...

Visualizing Adhesion Formation in Cells by Means of Advanced Spinning Disk-Total Internal Reflection Fluorescence Microscopy

利用先进的纺额圆盘全内反射荧光显微镜观察细胞粘附形成

In living cells, processes such as adhesion formation involve extensive structural changes in the plasma membrane and the cell interior. In order to ...