这项工作是支持的，部分由大学布法罗分校临床和转化研究中心翻译协会和水牛城罗斯威尔公园联盟基金会，启动资金的资助在布法罗大学，IT契合创意方案的试点研究计划的资助MKE，NIPA（C1515-1121-0003）及NRF授出的教育，科学和技术部（2012-0009249）。

1。深层思考模式光声膀胱造影系统（PAC） <ol><li>系统配置17，18 <ol><li> A Q-开关Nd:YAG激光（SLII-10;连续532纳米）泵的波长可调谐激光器（Surelite OPO PLUS连续波长调谐范围:680至2500纳米）。 </li><li>拍摄各激光的脉冲持续时间为〜5毫微秒，激光重复率是10赫兹。 </li><li>依赖于所使用的造影剂的光吸收峰的波长。如果亚甲基蓝作为造影剂，使用的光波长为667 nm，其中的峰值吸收。电浆黄金纳米结构可以有可调本地化的表面等离子体共振光谱区域，根据他们的物理和化学性质。此外，单壁碳纳米管的广泛吸收光谱波长选择可以提供多种选项。 </li><li>可调激光器的光被传递到一个球形的圆锥形晶状体吨即通过直角棱镜（PS9​​08，Thorlabs公司）。 </li><li>甲炸面圈状的光束图案生成球形的锥形透镜后的光穿过。自制的锥形透镜从BK7透镜和锥角为152°。透镜的直径为2.5厘米。 </li><li> 经由光学冷凝器，一个透明的丙烯酸系片材制成的炸面圈状的发散光束被重定向。的顶面和底面的直径分别为6.1和4.8厘米。冷凝器的厚度为2.5厘米。重定向的环形光束，形成一个环形的组织表面上一个黑暗的中心。 </li><li>利用一个小的水容器，以提高声耦合。水容器底部开有明显的细光学和声学透明的聚乙烯薄膜包裹。小动物的位置下，水的容器。 </li><li>生成的PA波检测由一个球形聚焦超声换能器（V308;奥林巴斯NDT; 5 MHz的中心频率），其被安装在中间的光学冷凝器。的换能器元件的直径和焦距分别为1.9和2.5厘米，分别。因此，换能器的F数为〜1.3。 </li><li>横向和轴向分辨率是590和144微米，分别。 </li><li>第一放大所检测到的PA波由一个宽带超声波脉冲发生器/接收器（5072PR奥林巴斯NDT; 35 MHz的带宽和射频增益59分贝），然后获取由示波器（TDS5054泰克）。 </li><li>取得一维时间分辨的图像（称为A线）PA的波的到达时间测量。目前，PA的波的速度被假定为在1,480米/秒，在整个图像采集。双（被称为B-扫描）和三维PA图像可以被收购，通过机械地移动阶段（XY6060，丹纳赫多佛），线性光栅扫描。 </li><li>的摄像的时间大约是25分钟为一个体积单波长PA图像与领域的视角（FOV）2.5倍2.4倍1.5厘米3的XYZ飞机。沿x方向，步长为0.2毫米，沿y方向与步长为0.4mm的60个样品获得125个样本。与50-MHz采样率的500个数据点，得到沿z方向。机械扫描控制的自制的NI LabVIEW软件系统。 </li><li>的体积数据是由使用MathWorks的MATLAB软件系统的最大振幅投影（地图）。 </li></ol></li><li>系统校准<ol><li>球形圆锥形晶状体后，确保光束模式是一个完美的环形状。如果没有正确生成的炸面圈状的波束图案，从皮肤表面PA的起源的信号占主导地位。因此，它是难以达到的深部组织的成像。 </li><li>线状光聚焦在水中应同轴对准超声波聚焦区域。那些被同轴对准，该系统遭受低信号对信噪比。 </li><li>在皮肤表面上照射的激光脉冲能量的变化范围为〜1 - 2毫焦耳/厘米2时，从680至1,000 nm的波长调谐时，分别。激光的脉冲能量比美国国家标准学会安全限值要低得多，为20〜80毫焦耳/厘米2以上的光谱区域，分别不同。 </li></ol></li></ol> （2） 在体内和体外成像过程<ol><li>动物准备<ol><li>使用女只SD大鼠，重量为200 - 250克所有PA成像实验。 </li><li>首先，麻醉大鼠腹腔注射氯胺酮（85毫克/公斤体重）和甲苯噻嗪（15毫克/千克）的混合物中。 </li><li>脱毛在腹部的毛发。 </li><li>大鼠定位一个定制的动物持有人之上。 </li><li>外套22号导管润滑剂以改善导管插入。 </li><li>保持导管Vertica的LLY超过尿道口。接下来，插入的导管的末端，水平，直到轮毂的导管插入尿道终于到达开口。 </li><li>尿液在膀胱经导管无效。 </li></ol></li><li> 在体内 PAC成像<ol><li>大鼠，位于动物保持器的顶部，下面的水容器中的PAC系统的位置。 </li><li>应用超声凝胶（SONOTECH）之间的动物的皮肤表面和塑料膜，以改善声学耦合。 </li><li>完全汽化的异氟醚（1升/分钟的氧和0.75％的异氟烷）的体内 PA成像的实验期间使用麻醉大鼠。 </li><li>获取控制PA图像注射造影剂前。 </li><li>引入亚甲基蓝（0.8微升/克，浓度为30 mM的体重），合金纳米笼（1.2微升/克的浓度为2纳米的体重），或单壁碳纳米管（0.8微升/克体的水溶液重量与的浓度为0.3μM） 经导管向膀胱。用1毫升注射器用22号导管。 </li><li>采集了一系列的PA图像。 </li></ol></li><li> 体外 PAC成像<ol><li>牺牲后大鼠体内 PAC成像，注入了过量的巴比妥。 </li><li>取出的两个主要器官，膀胱，肾脏，调查生物分布，在玻璃板上。 </li><li>玻璃板下面的水容器在PAC系统的位置。 </li><li>应用超声凝胶（SONOTECH）切除器官和塑料膜之间，以改善声学耦合。 </li><li>收购PA图像。 </li><li> Photoacoustically图像切除的器官从一个健康的老鼠作为对照。 </li></ol></li></ol>

<ol>
	<li>Riccabona, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cystography+in+infants+and+children:+a+critical+appraisal+of+the+many+forms+with+special+regard+to+voiding+cystourethrography.">Cystography in infants and children: a critical appraisal of the many forms with special regard to voiding cystourethrography.</a> Eur. Radiol. 12 (12), 2910-2918 (2002).</li><li>Khattar, N., Dorairajan, L. N., Kumar, S., Pal, B. C., Elangovan, S., Nayak, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Giant+obstructive+megaureter+causing+contralateral+ureteral+obstruction+and+hydronephrosis:+a+first-time+report.">Giant obstructive megaureter causing contralateral ureteral obstruction and hydronephrosis: a first-time report.</a> Urology. 74 (6), 1306-1308 (2009).</li><li>Lim, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vesicoureteral+reflux+and+urinary+tract+infection:+evolving+practices+and+current+controversies+in+pediatric+imaging.">Vesicoureteral reflux and urinary tract infection: evolving practices and current controversies in pediatric imaging.</a> AJR Am. J. Roentgenol. 192 (5), 1197-1208 (2009).</li><li>Scardapane, A., Pagliarulo, V., Ianora, A. A., Pagliarulo, A., Angelelli, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contrast-enhanced+multislice+pneumo-CT-cystography+in+the+evaluation+of+urinary+bladder+neoplasms.">Contrast-enhanced multislice pneumo-CT-cystography in the evaluation of urinary bladder neoplasms.</a> Eur. J. Radiol. 66 (2), 246-252 (2008).</li><li>Verpoorten, C., Buyse, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neurogenic+bladder:+medical+treatment.">The neurogenic bladder: medical treatment.</a> Pediatr. Nephrol. 23 (5), 717-725 (2008).</li><li>Ron, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Let's+not+relive+the+past:+a+review+of+cancer+risk+after+diagnostic+or+therapeutic+irradiation.">Let's not relive the past: a review of cancer risk after diagnostic or therapeutic irradiation.</a> Pediatr. Radiol. 32 (10), 739-744 (2002).</li><li>Berrington De Gonzalez, A., Darby, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Risk+of+cancer+from+diagnostic+X-rays:+estimates+for+the+UK+and+14+other+countries.">Risk of cancer from diagnostic X-rays: estimates for the UK and 14 other countries.</a> Lancet. 363 (9406), 345-351 (2004).</li><li>Kim, C., Favazza, C., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+photoacoustic+tomography+of+chemicals:+high-resolution+functional+and+molecular+optical+imaging+at+new+depths.">In vivo photoacoustic tomography of chemicals: high-resolution functional and molecular optical imaging at new depths.</a> Chem. Rev. 110 (5), 2756-2782 (2010).</li><li>Ke, H., Erpelding, T. N., Jankovic, L., Liu, C., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+characterization+of+an+integrated+ultrasound,+photoacoustic,+and+thermoacoustic+imaging+system.">Performance characterization of an integrated ultrasound, photoacoustic, and thermoacoustic imaging system.</a> J. Biomed. Opt. 17 (5), 056010 (2012).</li><li>Akers, W. J., Kim, C., Berezin, , 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=Noninvasive+Photoacoustic+and+Fluorescence+Sentinel+Lymph+Node+Identification+using+Dye-Loaded+Perfluorocarbon+Nanoparticles.">Noninvasive Photoacoustic and Fluorescence Sentinel Lymph Node Identification using Dye-Loaded Perfluorocarbon Nanoparticles.</a> Acs Nano. 5 (1), 173-182 (2011).</li><li>Jiao, S. L., Jiang, M. S., Hu, J., 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=Photoacoustic+ophthalmoscopy+for+in+vivo+retinal+imaging.">Photoacoustic ophthalmoscopy for in vivo retinal imaging.</a> Opt. Express. 18 (4), 3967-3972 (2010).</li><li>Kim, C., Cho, E. C., Chen, J., 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=In+vivo+molecular+photoacoustic+tomography+of+melanomas+targeted+by+bioconjugated+gold+nanocages.">In vivo molecular photoacoustic tomography of melanomas targeted by bioconjugated gold nanocages.</a> Acs Nano. 4 (8), 4559-4564 (2010).</li><li>Kim, C., Song, H. M., Cai, X., Yao, J., Wei, A., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+photoacoustic+mapping+of+lymphatic+systems+with+plasmon-resonant+nanostars.">In vivo photoacoustic mapping of lymphatic systems with plasmon-resonant nanostars.</a> J. Mater. Chem. 21 (9), 2841-2844 (2011).</li><li>Wang, X., Pang, Y., Ku, G., Xie, X., Stoica, G., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+laser-induced+photoacoustic+tomography+for+structural+and+functional+in+vivo+imaging+of+the+brain.">Noninvasive laser-induced photoacoustic tomography for structural and functional in vivo imaging of the brain.</a> Nat. Biotechnol. 21 (7), 803-806 (2003).</li><li>Xie, Z., Roberts, W., Carson, P., Liu, X., Tao, C., Wang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+bladder+microvasculature+with+high-resolution+photoacoustic+imaging.">Evaluation of bladder microvasculature with high-resolution photoacoustic imaging.</a> Opt. Lett. 36 (24), 4815-4817 (2011).</li><li>Zhang, H. F., Maslov, K., Stoica, G., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+photoacoustic+microscopy+for+high-resolution+and+noninvasive+in+vivo+imaging.">Functional photoacoustic microscopy for high-resolution and noninvasive in vivo imaging.</a> Nat. Biotechnol. 24 (7), 848-851 (2006).</li><li>Kim, C., Jeon, M., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonionizing+photoacoustic+cystography+in+vivo.">Nonionizing photoacoustic cystography in vivo.</a> Opt. Lett. 36 (18), 3599-3601 (2011).</li><li>Homan, K., Kim, S., Chen, Y. S., Wang, B., Mallidi, S., Emelianov, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prospects+of+molecular+photoacoustic+imaging+at+1064+nm+wavelength.">Prospects of molecular photoacoustic imaging at 1064 nm wavelength.</a> Opt. Lett. 35 (15), 2663-2665 (2010).</li><li>Chang, S. L., Shortliffe, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pediatric+urinary+tract+infections.">Pediatric urinary tract infections.</a> Pediatr. Clin. N. Am. 53 (3), 379 (2006).</li><li>Stratton, K. L., Pope, J. C., Adams, M. C., Brock, J. W., Thomas, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Implications of Ionizing Radiation in the Pediatric Urology. 183 (6), 2137-2142 (2010).</li><li>Ermilov, S. A., Khamapirad, T., Conjusteau, 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=Laser+optoacoustic+imaging+system+for+detection+of+breast+cancer.">Laser optoacoustic imaging system for detection of breast cancer.</a> J. Biomed. Opt. 14 (2), 024007 (2009).</li><li>Erpelding, T. N., Kim, C., Pramanik, M., 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=Sentinel+lymph+nodes+in+the+rat:+noninvasive+photoacoustic+and+US+imaging+with+a+clinical+US+system.">Sentinel lymph nodes in the rat: noninvasive photoacoustic and US imaging with a clinical US system.</a> Radiology. 256 (1), 102-110 (2010).</li><li>Kim, C., Erpelding, T. N., Jankovic, L., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performance+benchmarks+of+an+array-based+hand-held+photoacoustic+probe+adapted+from+a+clinical+ultrasound+system+for+non-invasive+sentinel+lymph+node+imaging.">Performance benchmarks of an array-based hand-held photoacoustic probe adapted from a clinical ultrasound system for non-invasive sentinel lymph node imaging.</a> Philos. Transact. A. Math Phys. Eng. Sci. 369 (1955), 4644-4650 (1955).</li><li>Kim, C., Song, K. H., Gao, F., Wang, L. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sentinel+lymph+nodes+and+lymphatic+vessels:+noninvasive+dual-modality+in+vivo+mapping+by+using+indocyanine+green+in+rats--volumetric+spectroscopic+photoacoustic+imaging+and+planar+fluorescence+imaging.">Sentinel lymph nodes and lymphatic vessels: noninvasive dual-modality in vivo mapping by using indocyanine green in rats--volumetric spectroscopic photoacoustic imaging and planar fluorescence imaging.</a> Radiology. 255 (2), 442-450 (2010).</li><li>Kruger, R. A., Kiser, W. L., Reinecke, D. R., Kruger, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermoacoustic+computed+tomography+using+a+conventional+linear+transducer+array.">Thermoacoustic computed tomography using a conventional linear transducer array.</a> Medical Physics. 30 (5), 856-860 (2003).</li><li>Kruger, R. A., Lam, R. B., Reinecke, D. R., Del Rio, S. P., Doyle, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photoacoustic+angiography+of+the+breast.">Photoacoustic angiography of the breast.</a> Med. Phys. 37 (11), 6096-6100 (2010).</li><li>Manohar, S., Kharine, A., Van Hespen, J. C., Steenbergen, W., Van Leeuwen, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Twente+Photoacoustic+Mammoscope:+system+overview+and+performance.">The Twente Photoacoustic Mammoscope: system overview and performance.</a> Phys. Med. Biol. 50 (11), 2543-2557 (2005).</li></ol>

在所有的动物实验符合纽约州立大学布法罗机构动物护理和使用委员会。 什么都没有透露。

总之，我们已经证明电离PAC使用无毒的光吸收剂在大鼠模型体内的可能性。我们已经成功地拍摄到大鼠膀胱充满了非电离和非侵入性的PAC系统使用光吸收剂。在我们的方法中两个关键的安全问题已经得到解决:（1）使用非电离辐射的cystographic应用和（2）造影剂，在体内无蓄积。 我们的临床权益包括监测膀胱输尿管返流（VUR）在儿科患者。在美国约3％的儿童都受到尿路?...

X射线膀胱造影1的成像过程中，以确定有关的膀胱疾病，如膀胱癌，膀胱输尿管回流，输尿管阻塞，神经原性膀胱， 等 。2-5通常情况下，尿空隙和一个无线电不透明剂，通过注入导尿管。然后，透视X射线图像采集划定尿膀胱。然而，关键的安全问题是，此过程中使用有害的电离辐射。累积的患癌症的风险的百分比75岁时由于诊断X射线范围从0.6至1.8％。6此外，致癌性的威胁是在儿童患者中具有重要意义。英国的一项研究表明，在9个主要内脏器官，平均每年诊断X射线辐射剂量最高的女性儿童的膀胱小于4和第二最高的男性儿童小于4 7这表明膀胱癌的风险。在儿科患者中最重要的。 Alt键霍夫儿科放射科医师努力以减轻辐射的暴露率，合理实现，电离辐射可低至不能完全排除。因此，限制创建完全辐射，敏感，具有成本效益，和高分辨率成像方式与非放射性造影剂在膀胱造影需要。 近日，光声断层成像（PAT）已成为首屈一指的生物医学成像方式，因为PAT可以提供强大的光学吸收的对比和高超声波在生物组织中的空间分辨率。8 PAT的原理是，超声波引起的热弹性膨胀的目标然后由光吸收。通过检测时间分辨的声波通过的介质，二维或三维的光声（PA）的图像形成。由于超声（US）要少得多组织中散射光（通常为两个或三个数量级），相比的PAT成像深度可达〜8厘米组织的空间分辨率，同时保持以1/200的成像深度。9的主要优点PAT为cystographic应用包括:（1）PAT是完全不受电离辐射。 （2）ClinicalUSimaging系统可以很容易地适合于提供双模式的PA和美国的成像能力。因此，双模式的PA /美国成像系统可以比较便携，成本效益，快速，这是临床快速翻译的关键标准。使用内源性和外源性对比，PAT提供高分辨率的形态，功能，组织和分子成像研究肿瘤病理生理学，脑血流动力学变化，内脏器官，眼科，血管造影等 。10-16 在这篇文章中，我们展示了非电离光声膀胱造影实验协议（PAC）的近红外（NIR）光吸收剂（ 即亚甲基蓝，去LD纳米笼，或单壁碳纳米管）为无毒光学浑浊示踪剂。大鼠膀胱充满了造影剂photoacoustically光谱体内划定的。没有代理人坚持积累的大鼠在膀胱和肾脏。因此，这可能是由于剂积累的长期毒性可以被排除在外。这一结果意味着PAC相结合的光吸收剂可以是一个潜在真正无害的cystographic的方式为儿童患者。系统配置，系统调整，并在体内 / 体外成像程序在这篇文章中讨论。

<table border="1">
	<tbody>
		<tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Q-switched Nd:YAG laser</td>
			<td>Continuum</td>
			<td>SLII-10</td>
			<td>pump laser</td>
		</tr>
		<tr>
			<td>OPO laser</td>
			<td>Continuum</td>
			<td>Surelite OPO PLUS</td>
			<td>tunable laser</td>
		</tr>
		<tr>
			<td>Prisms</td>
			<td>Thorlabs</td>
			<td>PS908</td>
			<td>light deliver</td>
		</tr>
		<tr>
			<td>Ultrasound transducer</td>
			<td>Olympus NDT</td>
			<td>V308</td>
			<td>5 MHz</td>
		</tr>
		<tr>
			<td>Ultraoundpulser/receiver</td>
			<td>Olympus NDT</td>
			<td>5072PR</td>
			<td>amplifier</td>
		</tr>
		<tr>
			<td>Oscilloscope</td>
			<td>Tektronix</td>
			<td>TDS5054</td>
			<td>data acquisition</td>
		</tr>
		<tr>
			<td>Scanning stage</td>
			<td>Danaher Dover</td>
			<td>XY6060</td>
			<td>raster scanning</td>
		</tr>
		<tr>
			<td>Methylene blue</td>
			<td>Sigma-Aldrich</td>
			<td>M9140-25G</td>
			<td>contrast agent</td>
		</tr>
		<tr>
			<td>Rats</td>
			<td>Harlan</td>
			<td>Spague-Dawley</td>
			<td>animal subject</td>
		</tr>
		<tr>
			<td>Isoflourane vaporizer</td>
			<td>Euthanex</td>
			<td>EZ-155</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Ultrasound gel</td>
			<td>Sonotech</td>
			<td>Clear Image singles</td>
			<td>acoustic coupling</td>
		</tr>
	</tbody>
</table>

photoacoustic cystography

常规小儿膀胱造影诊断X射线，这是基于使用不透射线的染料，遭受电离辐射的有害使用。由于暴露于辐射的儿童的膀胱癌的风险比许多其他癌症更重要。在这里，我们展示了非电离和非侵入性光声成像（PA）的膀胱，称为光声膀胱造影（PAC）的可行性，利用近红外（NIR）光吸收剂（ 即亚甲基蓝，电浆黄金纳米结构，单壁碳纳米管）的光学混浊示踪剂。我们已经成功地成像大鼠膀胱充满了用暗视场共焦PAC系统的光吸收剂。大鼠膀胱经尿道注射造影剂后，取得显著PA信号增强photoacoustically可视化。积累验证由光谱PA成像。此外，通过只用一个激光脉冲能量小于1兆焦耳/厘米2（1/20的安全限度的），我们目前的成像系统可以映射在体内在生物组织中的深度超过1 cm的亚甲基蓝填充大鼠膀胱。无论是在体内和体外 PA成像效果验证，造影剂通过小便自然排出。因此，不存在关切长期毒性剂的积累，这将有利于临床翻译。

图1显示了在体内非电离的，非侵入性的PAC:使用光学混浊的亚甲基蓝（MB）。的控制PA的图像，得到在667 nm处，在峰值光吸收MB（ 图1A）。虽然视场中的血管清晰可见，膀胱是不可见的，因为在此波长是光学透明的。 图1B中所示，膀胱清楚地表明在PA获取的图像在0.2小时后喷射甲基溴。要确认的积累MB在膀胱中，我们使用两个光波长（667和850纳米）来区分...

Watch this Scientific Journal Video about Photoacoustic Cystography at JoVE.com

Photoacoustic Cystography

光声膀胱造影（PAC），膀胱，辐射敏感的内部器官在儿科患者中，映射，而无需使用任何电离辐射或有毒造影剂具有很大的潜力。在这里，我们演示了如何使用一个注射的光学不透明示踪剂在大鼠尿膀胱映射PAC<em&gt;在体内</em&gt;。

光声膀胱造影

Conventional pediatric cystography, which is based on diagnostic X-ray using a radio-opaque dye, suffers from the use of harmful ionizing radiation. The ...

Research

JoVE Journal

Bioengineering

13.2K 次观看.  University at Buffalo, The State University of New York. 光声膀胱造影（PAC），膀胱，辐射敏感的内部器官在儿科患者中，映射，而无需使用任何电离辐射或有毒造影剂具有很大的潜力。在这里，我们演示了如何使用一个注射的光学不透明示踪剂在大鼠尿膀胱映射PAC在体内。

视频: Photoacoustic Cystography

Three-dimensional Optical-resolution Photoacoustic Microscopy

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As the mainstays of in vivo three-dimensional (3-D) optical microscopy, single-/multi-photon fluorescence microscopy and optical coherence tomography (OCT) have demonstrated their extraordinary sensitivities to fluorescence and optical scattering contrasts, respectively. However, the optical absorption contrast of biological tissues, which encodes essential physiological/pathological information, has not yet been assessable. 
The emergence of biomedical photoacoustics has led to a new branch of optical microscopy optical-resolution photoacoustic microscopy (OR-PAM)1, where the optical irradiation is focused to the diffraction limit to achieve cellular1 or even subcellular2 level lateral resolution. As a valuable complement to existing optical microscopy technologies, OR-PAM brings in at least two novelties. First and most importantly, OR-PAM detects optical absorption contrasts with extraordinary sensitivity (i.e., 100%). Combining OR-PAM with fluorescence microscopy3 or with optical-scattering-based OCT4 (or with both) provides comprehensive optical properties of biological tissues. Second, OR-PAM encodes optical absorption into acoustic waves, in contrast to the pure optical processes in fluorescence microscopy and OCT, and provides background-free detection. The acoustic detection in OR-PAM mitigates the impacts of optical scattering on signal degradation and naturally eliminates possible interferences (i.e., crosstalks) between excitation and detection, which is a common problem in fluorescence microscopy due to the overlap between the excitation and fluorescence spectra. 
Unique for optical absorption imaging, OR-PAM has demonstrated broad biomedical applications since its invention, including, but not limited to, neurology5, 6, ophthalmology7, 8, vascular biology9, and dermatology10. In this video, we teach the system configuration and alignment of OR-PAM as well as the experimental procedures for in vivo functional microvascular imaging.

Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technology capable of imaging optical absorption contrasts in vivo with cellular resolution and sensitivity. Here, we provide a visualized instruction on the experimental protocols of OR-PAM, including system configuration, system alignment, typical in vivo experimental procedures, and functional imaging schemes.

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

Detection and Isolation of Circulating Melanoma Cells using Photoacoustic Flowmetry

Circulating tumor cells (CTCs) are those cells that have separated from a macroscopic tumor and spread through the blood and lymph systems to seed secondary tumors1,2,3. CTCs are indicators of metastatic disease and their detection in blood samples may be used to diagnose cancer and monitor a patient&prime;s response to therapy. Since CTCs are rare, comprising about one tumor cell among billions of normal blood cells in advanced cancer patients, their detection and enumeration is a difficult task. We exploit the presence of pigment in most melanoma cells to generate photoacoustic, or laser induced ultrasonic waves in a custom flow cytometer for detection of circulating melanoma cells (CMCs)4,5. This process entails separating a whole blood sample using centrifugation and obtaining the white blood cell layer. If present in whole blood, CMCs will separate with the white blood cells due to similar density. These cells are resuspended in phosphate buffered saline (PBS) and introduced into the flowmeter. Rather than a continuous flow of the blood cell suspension, we induced two phase flow in order to capture these cells for further study. In two phase flow, two immiscible liquids in a microfluidic system meet at a junction and form alternating slugs of liquid6,7. PBS suspended white blood cells and air form microliter slugs that are sequentially irradiated with laser light. The addition of a surfactant to the liquid phase allows uniform slug formation and the user can create different sized slugs by altering the flow rates of the two phases. Slugs of air and slugs of PBS with white blood cells contain no light absorbers and hence, do not produce photoacoustic waves. However, slugs of white blood cells that contain even single CMCs absorb laser light and produce high frequency acoustic waves. These slugs that generate photoacoustic waves are sequestered and collected for cytochemical staining for verification of CMCs.

We have developed a flow cytometer using laser induced ultrasound to detect circulating melanoma cells as an early indicator of metastatic disease.

使用光声血流循环黑色素瘤细胞的检测和隔离

Circulating tumor cells (CTCs) are those cells that have separated from a macroscopic tumor and spread through the blood and lymph systems to seed ...

Integrated Photoacoustic Ophthalmoscopy and Spectral-domain Optical Coherence Tomography

Both the clinical diagnosis and fundamental investigation of major ocular diseases greatly benefit from various non-invasive ophthalmic imaging technologies. Existing retinal imaging modalities, such as fundus photography1, confocal scanning laser ophthalmoscopy (cSLO)2, and optical coherence tomography (OCT)3, have significant contributions in monitoring disease onsets and progressions, and developing new therapeutic strategies. However, they predominantly rely on the back-reflected photons from the retina. As a consequence, the optical absorption properties of the retina, which are usually strongly associated with retinal pathophysiology status, are inaccessible by the traditional imaging technologies.
Photoacoustic ophthalmoscopy (PAOM) is an emerging retinal imaging modality that permits the detection of the optical absorption contrasts in the eye with a high sensitivity4-7 . In PAOM nanosecond laser pulses are delivered through the pupil and scanned across the posterior eye to induce photoacoustic (PA) signals, which are detected by an unfocused ultrasonic transducer attached to the eyelid. Because of the strong optical absorption of hemoglobin and melanin, PAOM is capable of non-invasively imaging the retinal and choroidal vasculatures, and the retinal pigment epithelium (RPE) melanin at high contrasts 6,7. More importantly, based on the well-developed spectroscopic photoacoustic imaging5,8 , PAOM has the potential to map the hemoglobin oxygen saturation in retinal vessels, which can be critical in studying the physiology and pathology of several blinding diseases 9 such as diabetic retinopathy and neovascular age-related macular degeneration.
Moreover, being the only existing optical-absorption-based ophthalmic imaging modality, PAOM can be integrated with well-established clinical ophthalmic imaging techniques to achieve more comprehensive anatomic and functional evaluations of the eye based on multiple optical contrasts6,10 . In this work, we integrate PAOM and spectral-domain OCT (SD-OCT) for simultaneously in vivo retinal imaging of rat, where both optical absorption and scattering properties of the retina are revealed. The system configuration, system alignment and imaging acquisition are presented.

Photoacoustic ophthalmology (PAOM), an optical-absorption-based imaging modality, provides the complementary evaluation of the retina to the currently available ophthalmic imaging technologies. We report the using of PAOM integrated with spectral-domain optical coherence tomography (SD-OCT) for simultaneous multimodal retinal imaging in rats.

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 ...

Preparation and Photoacoustic Analysis of Cellular Vehicles Containing Gold Nanorods

Gold nanorods are attractive for a range of biomedical applications, such as the photothermal ablation and the photoacoustic imaging of cancer, thanks to their intense optical absorbance in the near-infrared window, low cytotoxicity and potential to home into tumors. However, their delivery to tumors still remains an issue. An innovative approach consists of the exploitation of the tropism of tumor-associated macrophages that may be loaded with gold nanorods in vitro. Here, we describe the preparation and the photoacoustic inspection of cellular vehicles containing gold nanorods. PEGylated gold nanorods are modified with quaternary ammonium compounds, in order to achieve a cationic profile. On contact with murine macrophages in ordinary Petri dishes, these particles are found to undergo massive uptake into endocytic vesicles. Then these cells are embedded in biopolymeric hydrogels, which are used to verify that the stability of photoacoustic conversion of the particles is retained in their inclusion into cellular vehicles. We are confident that these results may provide new inspiration for the development of novel strategies to deliver plasmonic particles to tumors.

We show the preparation and address the feasibility of cellular vehicles containing gold nanorods for the photoacoustic imaging of cancer.

含有金纳米棒蜂窝车辆的制备及光声分析

Gold nanorods are attractive for a range of biomedical applications, such as the photothermal ablation and the photoacoustic imaging of cancer, thanks to ...

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

Photoacoustic microscopy (PAM) is a fast-growing invivo imaging modality that combines both optics and ultrasound, providing penetration beyond the optical mean free path (~1 mm in skin) with high resolution. By combining optical absorption contrast with the high spatial resolution of ultrasound in a single modality, this technique can penetrate deep tissues. Photoacoustic microscopy systems can have either a low acoustic resolution and probe deeply or a high optical resolution and probe shallowly. It is challenging to achieve high spatial resolution and large depth penetration with a single system. This work presents an AR-OR-PAM system capable of both high-resolution imaging at shallow depths and low-resolution deep-tissue imaging of the same sample in vivo. A lateral resolution of 4 &#181;m with 1.4 mm imaging depth using optical focusing and a lateral resolution of 45 &#181;m with 7.8 mm imaging depth using acoustic focusing were successfully demonstrated using the combined system. Here,&#160;in vivo&#160;small-animal blood vasculature imaging is performed to demonstrate its biological imaging capability.

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

Here a switchable acoustic resolution (AR) and optical resolution (OR) photoacoustic microscopy (AR-OR-PAM) system capable of both high resolution imaging at shallow depth and low resolution deep tissue imaging on the same sample in vivo is demonstrated.

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 ...

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

In vivo small-animal imaging has an important role to play in preclinical studies. Photoacoustic tomography (PAT) is an emerging hybrid imaging modality that shows great potential for both preclinical and clinical applications. Conventional optical parametric oscillator-based PAT (OPO-PAT) systems are bulky and expensive and cannot provide high-speed imaging. Recently, pulsed-laser diodes (PLDs) have been successfully demonstrated as an alternative excitation source for PAT. Pulsed-laser diode PAT (PLD-PAT) has been successfully demonstrated for high-speed imaging on photoacoustic phantoms and biological tissues. This work provides a visualized experimental protocol for in vivo brain imaging using PLD-PAT. The protocol includes the compact PLD-PAT system configuration and its description, animal preparation for brain imaging, and a typical experimental procedure for 2D cross-sectional rat brain imaging. The PLD-PAT system is compact and cost-effective and can provide high-speed, high-quality imaging. Brain images collected in vivo at various scan speeds are presented.

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

A compact pulsed-laser diode-based photoacoustic tomography (PLD-PAT) system for high-speed in vivo brain imaging in small animals is demonstrated.

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

Translation of photoacoustic imaging into the clinic is a major challenge. Handheld real-time clinical photoacoustic imaging systems are very rare. Here, we report a combined photoacoustic and clinical ultrasound imaging system by integrating an ultrasound probe with light delivery for small animal imaging. We demonstrate this by showing sentinel lymph node imaging in small animals along with minimally invasive real-time needle guidance. A clinical ultrasound platform with access to raw channel data allows the integration of photoacoustic imaging leading to a handheld real-time clinical photoacoustic imaging system. Methylene blue was used for sentinel lymph node imaging at 675 nm wavelength. Additionally, needle guidance with dual modal ultrasound and photoacoustic imaging was shown using the imaging system. Depth imaging of up to 1.5 cm was demonstrated with a 10 Hz laser at a photoacoustic imaging frame rate of 5 frames per second.

A clinical handheld photoacoustic imaging system will be demonstrated 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, ...

Pulsed Laser Diode-Based Desktop Photoacoustic Tomography for Monitoring Wash-In and Wash-Out of Dye in Rat Cortical Vasculature

Photoacoustic (PA) tomography (PAT) imaging is an emerging biomedical imaging modality useful in various preclinical and clinical applications. Custom-made circular ring array-based transducers and conventional bulky Nd:YAG/OPO lasers inhibit translation of the PAT system to clinics. Ultra-compact pulsed laser diodes (PLDs) are currently being used as an alternative source of near-infrared excitation for PA imaging. High-speed dynamic in vivo imaging has been demonstrated using a compact PLD-based desktop PAT system (PLD-PAT). A visualized experimental protocol using the desktop PLD-PAT system is provided in this work for dynamic in vivo brain imaging. The protocol describes the desktop PLD-PAT system configuration, preparation of animal for brain vascular imaging, and procedure for dynamic visualization of indocyanine green (ICG) dye uptake and clearance process in rat cortical vasculature.&#160;

A compact pulsed laser diode-based desktop photoacoustic tomography (PLD-PAT) system is demonstrated for high-speed dynamic in vivo imaging of small animal cortical vasculature.

基于脉冲激光二极管的桌面光声层析成像在大鼠皮质血管内监测染料的洗入和洗脱

Photoacoustic (PA) tomography (PAT) imaging is an emerging biomedical imaging modality useful in various preclinical and clinical applications. ...

Ovarian Cancer Detection Using Photoacoustic Flow Cytometry

Many studies suggest that the enumeration of circulating tumor cells (CTCs) may show promise as a prognostic tool for ovarian cancer. Current strategies for the detection of CTCs include flow cytometry, microfluidic devices, and real-time polymerase chain reaction (RT-PCR). Despite recent advances, methods for the detection of early ovarian cancer metastasis still lack the sensitivity and specificity required for clinical translation. Here, a novel method is presented for the detection of ovarian circulating tumor cells by photoacoustic flow cytometry (PAFC) utilizing a custom three dimensional (3D) printed system, including a flow chamber and syringe pump. This method utilizes folic acid-capped copper sulfide nanoparticles (FA-CuS NPs) to target SKOV-3 ovarian cancer cells by PAFC. This work demonstrates the affinity of these contrast agents for ovarian cancer cells. The results show NP characterization, PAFC detection, and NP uptake by fluorescence microscopy, thus demonstrating the potential of this novel system to detect ovarian CTCs at physiologically relevant concentrations.

A protocol is presented to detect circulating ovarian tumor cells utilizing a custom-made photoacoustic flow system and targeted folic acid-capped copper sulfide nanoparticles.

使用光声流细胞测定卵巢癌检测

Many studies suggest that the enumeration of circulating tumor cells (CTCs) may show promise as a prognostic tool for ovarian cancer. Current strategies ...

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

Imaging of vascular networks on small animals has played an important role in basic biomedical research. Photoacoustic imaging technology has great potential for application in the imageology of small animals. The wide-field photoacoustic imaging of small animals can provide images with high spatiotemporal resolution, deep penetration, and multiple contrasts. Also, the real-time photoacoustic imaging system is desirable to observe the hemodynamic activities of small-animal vasculature, which can be used to research the dynamic monitoring of small-animal physiological features. Here, a dual-raster-scanning photoacoustic imager is presented, featuring a switchable double-mode imaging function. The wide-field imaging is driven by a two-dimensional motorized translation stage, while the real-time imaging is realized with galvanometers. By setting different parameters and imaging modes, in vivo visualization of small-animal vascular network can be performed. The real-time imaging can be used to observe pulse change and blood flow change of drug-induced, etc. The wide-field imaging can be used to track the growth change of tumor vasculature. These are easy to be adopted in various areas of basic biomedicine research.

A dual raster-scanning photoacoustic imager was designed, which integrated wide-field imaging and real-time imaging.

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 ...