这项工作得到国家科学基金会职业奖的支持。CBET-1254767 和国家过敏和传染性疾病研究所授予 no。R01 AI104960。我们感激地承认帕特里克格里芬和丹为他们的协助与定性测量和得克萨斯 A & M 心血管病理学实验室为微 CT 成像。

1. 基体材料性能的选择和验证
<ol><li>在启动幻像制造过程 (图 1) 之前, 请找出成像波长对感兴趣的生物组织的吸收和减少散射系数。初步估计数可能在引用35、36中找到。但是, 可能需要对光学系数进行验证。</li>
	<li>使用用于吸收系数的查找表、µa和减少的散射系数、µs'、488、535、632和 775 nm 波长 (表第1-4和图 2–3), 选择印度墨水的浓度和二氧化钛 (2), 近似于所需的光学性质。这些食谱是特定的幽灵制造的。由于这些表提供了离散波长的实验数据, 所以对于特定的应用, 可能需要对配方进行优化。</li>
	<li>
制作所选配方的烷板, 以确认其光学性能。
	<ol>
<li>用10:1 的重量比的树脂, 以固化剂, 倒入配料杯按以下顺序: 甲基丙烯酸树脂,2, 印度墨水, 丙烯酸酯固化剂。 注: 在这里, 我们测试两个食谱: 1) 2 毫克2 + 3.5 µL 印度墨水每 g 和 2) 1 毫克的2 + 10 µL 印度墨水每 g。对于每种配方, 4.5 克和0.45 克的甲基丙烯酸酯固化剂与相应数量的光学粒子一起使用。</li>
		<li>混合在一个速度混合器 (参见材料表) 六十年代。如果2粒子粘在混合杯上 (可能与高浓度的钛的2), 手工混合, 以去除从杯底的颗粒, 并混合在混合器为另一个三十年代。</li>
		<li>将混合物倒入水井或培养皿中, 使混合物的薄 (0.1-1 毫米) 板。</li>
		<li>把这些石板加在一个气密的负压室里, 然后把它放在一个预热的烤箱里, 在80摄氏度处放10分钟, 然后放凉。</li>
		<li>从容器中取出冷却的聚合物板。修剪边缘, 离开平坦, 均匀的板。用卡尺测量板坯的厚度。</li>
	</ol>
</li>
	<li>
用积分球面测量板的透射率 (T) 和反射率 (R)。其他详细信息和说明可以在反向添加加倍 (内部审计) 手册37中找到。
	<ol>
<li>打开集成球体设置的光源和光谱仪。检查系统的对准, 以确保一个小的, 准直光束集中在进入和退出端口的积分球。</li>
		<li>校准积分球系统。<ol>
<li>关闭源, 将积分球体的出口端口盖上, 并记录三暗光谱。</li>
			<li>重新打开源代码, 以获得与出口端口上限和入口端口空的传输参考。记录三光谱。</li>
			<li>使用反射率标准获得反射率参考测量。将每个标准放在球体的出口端口。记录每个反射率标准的三光谱。</li>
		</ol>
</li>
		<li>测量板的透射率。在出口端口上盖上盖子, 将板置于积分球体的入口端, 用于传输测量。记录三光谱。</li>
		<li>测量板的反射率。卸下出口端盖, 并将底板放在出口端口上以进行反射测量。记录三光谱。</li>
	</ol>
</li>
	<li>
使用内部网络软件确定光学性能。本软件的完整教程可以在 "内部网络手册" 中找到, 软件下载37,38。
	<ol>
<li>平均每项测量获得的三光谱。</li>
		<li>使用内部网络手册37中的等式, 将这些测量转换为 R 和 T 值。如有必要, 通过降低频谱采样率来压缩文件。</li>
		<li>准备输入. rxt 文件 (补充材料 1), 使其具有波长、反射率、透射率和样本厚度, 如内部审计手册37中所述。使用命令提示符 (Windows os) 或终端 (Mac os), 导航到正确的路径。键入 "内部审计" 输入文件名 "" 以运行内部网络。该软件将生成具有估计光学属性的输出文本文件。</li>
	</ol>
</li>
	<li>如果光学属性不在所需值的可接受范围 (~ 15%) 之内, 请相应地修改配方并重复步骤1.3–1.5。</li>
</ol>2. 可溶性3D 印刷内部结构的制备
<ol><li>采用计算机辅助设计 (CAD) 软件设计内部结构。将结构实体模型转换为光固化成形文件, 以便在3D 打印机上进行制作。如果可用, 分割的 CT 扫描也可以转换成一个光固化成形文件, 而不是绘制一个实体模型的内部结构。 注意: 此处使用的分形树结构的 CAD 文件在补充材料 2中提供。本文所用的打印机是一种挤出式打印机, 因此该零件设计为具有断距支撑材料。</li>
	<li>选择用于打印的可溶性材料, 如聚乙烯醇 (PVA) 或高冲击聚苯乙烯 (臀部) (请参阅材料表)。在这种可溶性材料中打印固体模型。</li>
	<li>当印刷零件充分冷却, 打破, 溶解, 或机器的支持材料的印刷部分。文件或沙子关闭任何大瑕疵。</li>
	<li>
蒸气抛光印刷的部分, 以减少表面粗糙度。
	<ol>
<li>与印刷部分担保的副, 钻一个通孔与间隙的薄钢或镍钛丝在底部的印刷部分。</li>
		<li>螺纹不锈钢或镍钛丝通过孔。弯曲两端的导线和钩在一起。这将允许部分完全沉浸在丙酮蒸气烧杯内。把电线和零件放在一边。</li>
		<li>填充一个大烧杯大约10% 满丙酮。将烧杯放在热板上, 加热到100摄氏度。注意: 在通风罩中执行此步骤, 以防止吸入丙酮蒸气。</li>
		<li>当丙酮蒸气凝结达到约一半的烧杯壁上时, 将环形导线与模拟气道挂在第二根导线上, 并在丙酮蒸气中悬浮 15–30 s. 确保打印部件不接触烧杯壁或彼此 (如果蒸气抛光一次多个部件)。</li>
		<li>移除打印部件, 并在空烧杯或容器上挂起。让部分干至少4小时。</li>
	</ol>
</li>
	<li>根据需要验证内部结构的尺寸是否符合 CAD 设计的公差。根据精度要求, 卡尺或3D 激光扫描仪可用于测量结构。</li>
</ol>3. 耐热模具的施工
注: 准备一个防泄漏, 耐热模具, 形成了一个人的幻影。选择一个模具几何最适合最终的幻影设计。这里介绍了一种可重用的矩形模具。
<ol><li>设计一个坚实的模具底座, 以3D 打印。该模具是为一个幽灵的设计为1.17 厘米 x 1.79 厘米的基础。该模具的底座有一个1毫米厚和5毫米深凹槽, 内部尺寸匹配的幻影基地。这使模具专用线, 并拆卸和重新使用模具。</li>
	<li>为模具打印一个底座, 具有足够的宽度, 以确保模具的专用线。</li>
	<li>将专用线放在模具底座的凹槽内。这里, 1 毫米厚的聚碳酸酯片被用作模具专用线。</li>
	<li>使用耐热胶带, 密封模具的边缘。在成型过程中, 所有边角和边缘都必须充分密封, 不要有气泡, 以防出现任何渗漏。</li>
	<li>在步骤3.4 中准备的模具内放置聚碳酸酯基板。该基板是相同的1毫米厚聚碳酸酯片作为模具壁板, 并给予幻影基一个平滑的表面, 不粗糙的3D 打印表面的模具底座。将完全干燥的蒸气抛光部分粘附到基板上。让胶水有足够的时间干。</li>
</ol>4. 聚合物幻影的制备
注: 使用在步骤1中确定的用于特定应用程序的批量矩阵材料的已验证配方。这里的协议提供了一个健康的小鼠肺组织幻影的步骤535毫微米与µs"40 cm-1和µ 2 cm-1。在制造过程中, 制造一个没有光学粒子的第二个幻影是有用的。
<ol><li>将9.1 克的聚合物树脂倒入塑料搅拌杯中。添加20毫克金红石2, 后跟35µl 印度墨水。最后将0.91 克固化剂添加到混合物的顶部。在步骤1.3.2 中遵循混合协议。</li>
	<li>将最终聚合物混合物倒入耐热模具中。</li>
	<li>将少量的混合物倒入一个单独的容器中, 以创建用于确认材料光学特性的聚合物板。确保足够的聚合物浇注到有至少100µm 厚度的板。</li>
	<li>将模拟气道模具和单独的板放在钟罐中进行脱气。开始真空过程。如果聚合物在模拟气道模具开始上升, 让空气回到钟罐爆裂的表面气泡, 然后开始拉空气。重复这个过程, 直到聚合物没有显着上升。这将采取5-10 分钟之间取决于多少空气被困在步骤4.2。一旦该市不再上涨, 继续向德加再加15分钟。</li>
	<li>脱气后, 慢慢让空气回到房间里。去除模拟气道幻影和聚合物板, 并放置在水平烤箱80°c 2 小时。</li>
	<li>从烤箱中取下幻影和石板, 让冷却20分钟. 用手术刀拆卸聚合物模, 不切割固化聚合物。将基板从模拟气道底座上折断。</li>
	<li>将幻像置于加热 (60 °c) ~ 0.5 米氢氧化钠 (氢氧化钠) 基槽内, 直到内部部分完全溶解为止。光学上清晰的参考幻像可以帮助确定内部元件的溶解时间。一旦内部结构被溶解, 把幻影从浴缸中取出, 在采取任何光学测量之前, 让它完全干燥 (~ 24 小时)。</li>
</ol>5. 幻影制造的验证
<ol><li>如果需要, 请使用高分辨率磁共振成像 (MRI) 或微计算机断层扫描 (CT) 成像来验证幻像几何。这些方法通过 < 400 µm39、40的轴向分辨率, 为混浊材料内的内部结构提供3D 验证。或者, 光学上清晰的参考幻影可以光学成像, 以验证打印部分完全溶解, 剩余的空隙是正确的几何。 注意: 我们已经验证了光学不透明幻影 (2 毫克的2 + 3.5 µl 印度墨水) 的内部几何, 并在北极星成像 (NSI) X50 上微 CT。在所有维度 (补充材料 3, 4) 中, 幻像被映像为20µm 分辨率。</li>
	<li>使用聚合物板和积分球体验证幻影的光学特性 (在步骤1.5–1.6 中描述)。</li>
</ol>

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Z., Chavan, A. V., Lin, D., Tsai, E. H. R., Webb, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+and+application+of+heterogeneous+printed+mouse+phantoms+for+whole+animal+optical+imaging.">Fabrication and application of heterogeneous printed mouse phantoms for whole animal optical imaging.</a> Appl Opt. 55 (2), 280-287 (2016).</li><li>Diep, P., 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=Three-dimensional+printed+optical+phantoms+with+customized+absorption+and+scattering+properties.">Three-dimensional printed optical phantoms with customized absorption and scattering properties.</a> Biomed Opt Express. 6 (11), 4212-4220 (2015).</li><li>Pogue, B. W., Patterson, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review+of+tissue+simulating+phantoms+for+optical+spectroscopy,+imaging+and+dosimetry.">Review of tissue simulating phantoms for optical spectroscopy, imaging and dosimetry.</a> Journal of Biomedical Optics. 11 (4), 041102-041116 (2006).</li><li>de Bruin, D. 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=Optical+phantoms+of+varying+geometry+based+on+thin+building+blocks+with+controlled+optical+properties.">Optical phantoms of varying geometry based on thin building blocks with controlled optical properties.</a> J Biomed Opt. 15 (2), 025001-025010 (2010).</li><li>Boyle, A. 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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fiber+optic+microendoscopy+for+preclinical+study+of+bacterial+infection+dynamics.">Fiber optic microendoscopy for preclinical study of bacterial infection dynamics.</a> Biomed Opt Express. 2 (5), 1121-1134 (2011).</li><li>Nooshabadi, F., 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=Intravital+fluorescence+excitation+in+whole-animal+optical+imaging.">Intravital fluorescence excitation in whole-animal optical imaging.</a> PLoS One. 11 (2), 0149932 (2016).</li><li>Nooshabadi, F., 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=Intravital+excitation+increases+detection+sensitivity+for+pulmonary+tuberculosis+by+whole-body+imaging+with+beta-lactamase+reporter+enzyme+fluorescence.">Intravital excitation increases detection sensitivity for pulmonary tuberculosis by whole-body imaging with beta-lactamase reporter enzyme fluorescence.</a> J Biophotonics. , (2016).</li><li>Duck, F. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Physical Properties of Tissue: A Comprehensive Reference Book. , (1990).</li><li>Tuchin, V. V., Tuchin, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Tissue optics: light scattering methods and instruments for medical diagnosis. 13, (2007).</li><li>Prahl, 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> Everything I think you should know about Inverse Adding-Doubling. , (2011).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Inverse Adding-Doubling v.3-9-12. , (2014).</li><li>Link, T. 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=A+Comparative+Study+of+Trabecular+Bone+Properties+in+the+Spine+and+Femur+Using+High+Resolution+MRI+and+CT.">A Comparative Study of Trabecular Bone Properties in the Spine and Femur Using High Resolution MRI and CT.</a> J Bone Miner Res. 13 (1), 122-132 (1998).</li><li>Batiste, D. 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=High-resolution+MRI+and+micro-CT+in+an+ex+vivo+rabbit+anterior+cruciate+ligament+transection+model+of+osteoarthritis.">High-resolution MRI and micro-CT in an ex vivo rabbit anterior cruciate ligament transection model of osteoarthritis.</a> Osteoarthr cartil. 12 (8), 614-626 (2004).</li><li>Greening, G. 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=Characterization+of+thin+poly(dimethylsiloxane)-based+tissue-simulating+phantoms+with+tunable+reduced+scattering+and+absorption+coefficients+at+visible+and+near-infrared+wavelengths.">Characterization of thin poly(dimethylsiloxane)-based tissue-simulating phantoms with tunable reduced scattering and absorption coefficients at visible and near-infrared wavelengths.</a> J Biomed Opt. 19 (11), 115002 (2014).</li><li>Meissner, S., Knels, L., Krueger, A., Koch, T., Koch, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+three-dimensional+optical+coherence+tomography+and+intravital+microscopy+for+imaging+subpleural+pulmonary+alveoli+in+isolated+rabbit+lungs.">Simultaneous three-dimensional optical coherence tomography and intravital microscopy for imaging subpleural pulmonary alveoli in isolated rabbit lungs.</a> J Biomed Opt. 14 (5), 054020 (2009).</li><li>Morris, A. 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作者没有什么可透露的。

我们已经展示了一种制造光学幽灵的方法, 用来代表小鼠肺, 用内部分支结构模拟内部空气组织界面。小鼠肺组织的光学特性是通过将独特的光学散射浓度和吸收粒子分布在 homogenously 内的聚合物来实现的。这些光学特性可以在不同状态 (即健康与病态组织) 的不同光谱范围内模拟生理值。光学特性取决于感兴趣的波长、基材和幻像内粒子的浓度。然而, 对于多个粒子, 散射和吸收之间的关系?...

组织幽灵广泛用于光学成像和光谱学仪器的系统表征和校准, 包括包含超声波或核模式的综合系统1,2,3 ,4。幽灵为多种生物成像技术的系统表征和质量控制提供了受控光学环境。组织仿幽灵是预测系统性能和优化系统设计的重要工具, 在手头的生理任务;例如, 预测用于评估肿瘤边缘的光谱探针的探测深度5。幽灵的光学特性和结构设计可以被调谐来模拟使用该仪器的特定生理环境, 因此既允许可行性研究, 也可验证系统性能3, 6,7。在进入临床前或临床试验之前, 用逼真的光学幽灵验证成像系统的性能, 可以减少在体内研究期间出现故障或无法获取数据的风险。光学幽灵的重现性和稳定性使它们可定制光学技术的校准标准, 以监测内部和仪器间的变异性, 特别是在多中心临床试验中使用不同的仪器,运算符和环境条件8,9。
组织模拟幽灵也作为可调谐和可重现的物理模型, 用于验证理论光学模型。模拟有助于设计和优化在体内光学仪器, 同时减少对动物实验的需要10,11。光学模拟的发展和验证, 以准确地表示在体内环境可以由复杂的组织结构, 生物化学的内容, 以及目标或组织的位置在体内。不同学科之间的变异性使验证的理论模型具有挑战性的使用动物或人类的测量。聚合物光学组织幽灵允许通过提供已知和可再生的光学环境来验证理论模型, 研究光子迁移12,13,14,15。
在系统标定的目的, 固体光学幽灵可能包括一个单一的均匀板的固化聚合物与光学散射, 吸收, 或荧光调谐波长的兴趣。层状聚合物幽灵经常用于模拟上皮组织模型中组织光学特性的深度方差16,17。这些幻像结构是足够的上皮成像和建模, 因为组织结构是相当均匀, 通过每一层。然而, 更大的规模和更复杂的结构影响辐射运输在其他器官。为模拟皮下血管的光学环境, 建立了更复杂的幽灵的方法18,19甚至整个器官, 如膀胱20。由于空气组织界面的分支结构, 肺中光传输的建模提供了一个独特的问题;固体幻影不可能准确地在器官中复制辐射传输21。为了描述一种将复杂结构并入光学幻像的方法, 我们描述了一种方法, 用于创建一个表示气道三维 (3D) 宏观结构的内部可重现分形树 void (图 1)。
在过去的几十年中, 3D 打印已成为一个主要的方法, 快速原型医疗设备和模型22, 和光学组织幽灵也不例外。3D 印刷作为一种添加剂制造工具, 用于制作具有通道23、血管网络24和全身小动物模型25的光学幽灵。这些方法使用一个或两个具有独特光学性质的印刷材料。还开发了一些方法来调整印刷材料的光学特性, 以模拟一般的, 混浊的生物组织25,26。然而, 可实现光学性能的范围受到印刷材料的限制, 通常是聚合物, 如丙烯腈丁二烯苯乙烯 (ABS)26, 所以这种方法不适合所有的生物组织。烷 () 是一种光学上清晰的聚合物, 可以很容易地与散射和吸收粒子混合, 具有较高水平的调谐27,28。此外, 在为部署栓塞设备29、30的动脉瘤模型中, 还使用了用于建模的幽灵。这些幽灵还利用了一个可溶性的3D 打印部分, 但保持光学清晰, 以可视化设备部署。本文将该方法与调谐散射和吸收粒子的光学性质结合起来, 制备了小鼠肺组织和气道的初步模型。
虽然这里所呈现的幻影是特定于肺部, 这个过程可以应用到各种其他器官。3D. 幻影的内部结构的打印允许设计可以为任何目的和可打印的规模, 无论是血液或淋巴血管网络, 骨髓, 甚至心脏的四腔结构31。因为我们对肺的光学成像和建模有兴趣32,33,34, 我们选择使用四代分形树作为内部结构在聚合物幻影内复制。该结构的设计, 以近似的分支结构的气道和有突破支持材料的3D 印刷过程。如果不需要防断支架材料, 可以打印出更具解剖学上正确的气道。虽然这个特定的模型代表一个气道, 但幻影的内部结构并不需要保持物质的空隙。一旦周围的聚合物固化, 3D 印刷部分被溶解, 内部结构可以作为一个流动路径或作为一个二级模具的材料具有自己独特的吸收和散射特性。例如, 如果该协议的内部结构被设计成数字骨而不是气道, 则骨骼结构可以是3D 打印, 用与该手指的光学特性进行模压, 然后从幻影中溶解出来。然后, 可以用不同光学性质的混合材料填充该空隙。此外, 每个模具不限于一个单一的可溶性部分。手指的幻像可以被创造, 包括骨骼, 静脉, 动脉和一般的软组织层, 每个都有自己独特的光学特性。

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			<td height="42" style="height:42px;width:228px;">Dow Corning Sylgard 184 Silicone Encapsulant Clear 0.5 kg Kit</td>
			<td style="width:195px;">Ellsworth Adhesives</td>
			<td style="width:187px;">184 SIL ELAST KIT 0.5KG</td>
			<td style="width:259px;">Polydimethylsiloxane: polymer base for optical phantoms</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:228px;">White Rutile Titanium Dioxide powder</td>
			<td style="width:195px;">Atlantic Equipment Engineers</td>
			<td style="width:187px;">TI-602</td>
			<td style="width:259px;">Scattering particles for optical phantoms</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:228px;">Higgins Fountain Pen India Ink</td>
			<td style="width:195px;">Michaels Craft Stores</td>
			<td style="width:187px;">&#160;10015483</td>
			<td style="width:259px;">Absorbing particles for optical phantom</td>
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			<td height="21" style="height:21px;width:228px;">Heat Resistant tape</td>
			<td style="width:195px;">Uline</td>
			<td style="width:187px;">S-7595</td>
			<td style="width:259px;">Heat resistant tape for polymer molds</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:228px;">Fortus 360mc 3D printer</td>
			<td style="width:195px;">Stratasys</td>
			<td style="width:187px;">N/A</td>
			<td style="width:259px;">Able to switch build and support material with this model printer</td>
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		<tr height="63">
			<td height="63" style="height:63px;width:228px;">ABS Ivory Model Material</td>
			<td style="width:195px;">Stratasys</td>
			<td style="width:187px;">SDS-000001</td>
			<td style="width:259px;">Material for printing mold parts and/or using as support for printing internal structure&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:228px;">SR-30 Soluble Support</td>
			<td style="width:195px;">Stratasys</td>
			<td style="width:187px;">400638-0001</td>
			<td style="width:259px;">Base soluble support material for printing internal structure</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:228px;">Flacktek Speedmixer</td>
			<td style="width:195px;">Flacktek Inc.</td>
			<td style="width:187px;">DAC 150.1 FV</td>
			<td style="width:259px;">For efficient mixing of polymer and particles&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:228px;">Integrating sphere</td>
			<td style="width:195px;">Edmund Optics</td>
			<td style="width:187px;">58-585</td>
			<td style="width:259px;">For measuring optical properties</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:228px;">Polycarbonate build plates (1 mm)</td>
			<td style="width:195px;">Stratasys</td>
			<td style="width:187px;">N/A</td>
			<td style="width:259px;">Used polycarbonate build plates from Stratasys printer can also be used</td>
		</tr>
	</tbody>
</table>

fabrication and characterization of optical tissue phantoms containing macrostructure

新的光学成像技术的迅速发展取决于低成本、可定制和易于重现的标准的可用性。通过复制成像环境, 可以规避昂贵的动物实验来验证技术。预测和优化体内和前体成像技术的性能需要对与感兴趣的组织光学相似的样本进行测试。组织模仿光学幽灵为光学系统的评估、表征或校准提供了标准。均匀聚合物光学组织幽灵被广泛地用于模仿一个特定组织类型的光学性质在狭窄的光谱范围之内。层状组织, 如表皮和真皮, 可以通过简单地堆叠这些均质板幽灵来模拟。然而, 许多体内成像技术被应用于空间更复杂的组织, 如血管、呼吸道或组织缺损等三维结构会影响成像系统的性能。
本协议描述了一种组织模拟幻影的制作, 该幻像采用具有组织光学性质的材料的三维结构复杂度。查找表提供印度墨水和二氧化钛配方的光学吸收和散射目标。介绍了材料光学特性的表征和调优方法。本文详述的幻像制造有一个内部分支模拟气道空隙;然而, 该技术可以广泛应用于其他组织或器官结构。

为了演示幻像制作技术, 制作了小鼠肺组织幽灵, 以模拟 535 nm (表 5) 切除的健康和发炎的鼠肺组织的光学特性。这一波长的兴趣是 tdTomato 荧光蛋白的激发波长在重组报告菌株的分支杆菌在以前的研究33。用步骤1.4–1.5 中描述的方法得到了小鼠肺组织的光学测量。在德州一 & M 大学, 动物保育和使用委员会 (IACUC) 批准了对牲畜的使用。在 535 nm ?...

Watch this Scientific Journal Video about Fabrication and Characterization of Optical Tissue Phantoms Containing Macrostructure at JoVE.com

Fabrication and Characterization of Optical Tissue Phantoms Containing Macrostructure

光学组织幽灵是光学成像系统标定、表征和理论模型验证的基本工具。本文详细介绍了一种幻像制作方法, 包括组织光学特性和三维组织结构的复制。

含有宏观结构的光学组织幻影的制备与表征

The rapid development of new optical imaging techniques is dependent on the availability of low-cost, customizable, and easily reproducible standards. By ...

fabrication-characterization-optical-tissue-phantoms-containing

Research

JoVE Journal

Bioengineering

10.5K 次观看.  Texas A&M University. 光学组织幽灵是光学成像系统标定、表征和理论模型验证的基本工具。本文详细介绍了一种幻像制作方法, 包括组织光学特性和三维组织结构的复制。

视频: Fabrication and Characterization of Optical Tissue Phantoms Containing Macrostructure

Fabrication of Biologically Derived Injectable Materials for Myocardial Tissue Engineering

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications. Briefly, decellularized tissue is lyophilized, milled, enzymatically digested, and then brought to physiological pH. The lyophilization removes all water content from the tissue, resulting in dry ECM that can be ground into a fine powder with a small mill. After milling, the ECM powder is digested with pepsin to form an injectable matrix. After adjustment to pH 7.4, the liquid matrix material can be injected into the myocardium. Results of previous characterization assays have shown that matrix gels produced from decellularized pericardial and myocardial tissue retain native ECM components, including diverse proteins, peptides and glycosaminoglycans. Given the use of this material for tissue engineering, in vivo characterization is especially useful; here, a method for performing an intramural injection into the left ventricular (LV) free wall is presented as a means of analyzing the host response to the matrix gel in a small animal model. Access to the chest cavity is gained through the diaphragm and the injection is made slightly above the apex in the LV free wall. The biologically derived scaffold can be visualized by biotin-labeling before injection and then staining tissue sections with a horse radish peroxidase-conjugated neutravidin and visualizing via diaminobenzidine (DAB) staining. Analysis of the injection region can also be done with histological and immunohistochemical staining. In this way, the previously examined pericardial and myocardial matrix gels were shown to form fibrous, porous networks and promote vessel formation within the injection region.

Methods for preparing an injectable matrix gel from decellularized tissue and injecting it into rat myocardium in vivo are described.

心肌组织工程生物衍生的注射材料的制备

This protocol provides methods for the preparation of an injectable extracellular matrix (ECM) gel for myocardial tissue engineering applications.  ...

科研

生物工程

Fabrication of Micro-tissues using Modules of Collagen Gel Containing Cells

This protocol describes the fabrication of a type of micro-tissues called modules.  The module approach generates uniform, scalable and vascularized tissues.  The modules can be made of collagen as well as other gelable or crosslinkable materials.  They are approximately 2 mm in length and 0.7 mm in diameter upon fabrication but shrink in size with embedded cells or when the modules are coated with endothelial cells.  The modules individually are small enough that the embedded cells are within the diffusion limit of oxygen and other nutrients but modules can be packed together to form larger tissues that are perfusable.  These tissues are modular in construction because different cell types can be embedded in or coated on the modules before they are packed together to form complex tissues.  There are three main steps to making the modules: (1) neutralizing the collagen and embedding cells in it, (2) gelling the collagen in the tube and cutting the modules and (3) coating the modules with endothelial cells.

Creation of micro-tissues using cylindrical collagen gels, called modules, that contain embedded cells and which surface is coated with endothelial cells.

微组织使用胶原蛋白的模块的制备凝胶含有细胞

This protocol describes the fabrication of a type of micro-tissues called modules.  The module approach generates uniform, scalable and vascularized ...

Fabrication and Application of Rose Bengal-chitosan Films in Laser Tissue Repair

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two tissue edges1,2. These are then irradiated by a laser that is selectively absorbed by the RB. The resulting photochemical reactions supposedly crosslink the collagen fibers in the tissue with minimal heat production3. In this report, RB has been incorporated in thin chitosan films to fabricate a novel tissue adhesive that is laser-activated. Adhesive films, based on chitosan and containing ~0.1 wt% RB, are fabricated and bonded to calf intestine and rat tibial nerves by a solid state laser (&lambda;=532 nm, Fluence~110 J/cm2, spot size~0.5 cm). A single-column tensiometer, interfaced with a personal computer, is used to test the bonding strength. The RB-chitosan adhesive bonds firmly to the intestine with a strength of 15 &plusmn; 6 kPa, (n=30). The adhesion strength drops to 2 &plusmn; 2 kPa (n=30) when the laser is not applied to the adhesive. The anastomosis of tibial nerves can be also completed without the use of sutures. A novel chitosan adhesive has been fabricated that bonds photochemically to tissue and does not require sutures.

Sutures are usually needed to repair tissue during surgical procedures. However, their application can be problematic as they are invasive and may damage tissue. The fabrication and application methods of a novel tissue adhesive are here reported. This adhesive film is laser-activated and does not require the use of sutures.

激光对生物组织的修复孟加拉玫瑰红壳聚糖膜的制备及应用

Photochemical tissue bonding (PTB) is a sutureless technique for tissue repair, which is achieved by applying a solution of rose bengal (RB) between two ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

人体软骨组织制作采用三维喷墨打印技术

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Fabrication and Characterization of a Conformal Skin-like Electronic System for Quantitative, Cutaneous Wound Management

Recent advances in the development of electronic technologies and biomedical devices offer opportunities for non-invasive, quantitative assessment of cutaneous wound healing on the skin. Existing methods, however, still rely on visual inspections through various microscopic tools and devices that normally include high-cost, sophisticated systems and require well trained personnel for operation and data analysis. Here, we describe methods and protocols to fabricate a conformal, skin-like electronics system that enables conformal lamination to the skin surface near the wound tissues, which provides recording of high fidelity electrical signals such as skin temperature and thermal conductivity. The methods of device fabrication provide details of step-by-step preparation of the microelectronic system that is completely enclosed with elastomeric silicone materials to offer electrical isolation. The experimental study presents multifunctional, biocompatible, waterproof, reusable, and flexible/stretchable characteristics of the device for clinical applications. Protocols of clinical testing provide an overview and sequential process of cleaning, testing setup, system operation, and data acquisition with the skin-like electronics, gently mounted on hypersensitive, cutaneous wound and contralateral tissues on patients.

This article presents methods to fabricate and characterize a conformal, skin-like electronic system and protocols for the use in clinical applications, particularly on cutaneous wound management.

制作定量，皮肤伤口管理一个形皮肤般的电子系统与表征

Recent advances in the development of electronic technologies and biomedical devices offer opportunities for non-invasive, quantitative assessment of ...

Minced Tissue in Compressed Collagen: A Cell-containing Biotransplant for Single-staged Reconstructive Repair

Conventional techniques for cell expansion and transplantation of autologous cells for tissue engineering purposes can take place in specially equipped human cell culture facilities. These methods include isolation of cells in single cell suspension and several laborious and time-consuming events before transplantation back to the patient. Previous studies suggest that the body itself could be used as a bioreactor for cell expansion and regeneration of tissue in order to minimize ex vivo manipulations of tissues and cells before transplanting to the patient. The aim of this study was to demonstrate a method for tissue harvesting, isolation of continuous epithelium, mincing of the epithelium into small pieces and incorporating them into a three-layered biomaterial. The three-layered biomaterial then served as a delivery vehicle, to allow surgical handling, exchange of nutrition across the transplant, and a controlled degradation. The biomaterial consisted of two outer layers of collagen and a core of a mechanically stable and slowly degradable polymer. The minced epithelium was incorporated into one of the collagen layers before transplantation. By mincing the epithelial tissue into small pieces, the pieces could be spread and thereby the propagation of cells was stimulated. After the initial take of the transplants, cell expansion and reorganization would take place and extracellular matrix mature to allow ingrowth of capillaries and nerves and further maturation of the extracellular matrix. The technique minimizes ex vivo manipulations and allow cell harvesting, preparation of autograft, and transplantation to the patient as a simple one-stage intervention. In the future, tissue expansion could be initiated around a 3D mold inside the body itself, according to the specific needs of the patient. Additionally, the technique could be performed in an ordinary surgical setting without the need for sophisticated cell culturing facilities.

Tissue engineering often includes in vitro expansion in order to create autografts for tissue regeneration. In this study a method for tissue expansion, regeneration, and reconstruction in vivo was developed in order to minimize the processing of cells and biological materials outside the body.

在压缩的胶原组织糜:含细胞Biotransplant单上演重建修复

Conventional techniques for cell expansion and transplantation of autologous cells for tissue engineering purposes can take place in specially equipped ...

Fabrication of Extracellular Matrix-derived Foams and Microcarriers as Tissue-specific Cell Culture and Delivery Platforms

Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus of this article is on the methods for fabricating ECM-derived porous foams and microcarriers for use as biologically-relevant substrates in advanced 3D in vitro cell culture models or as pro-regenerative scaffolds and cell delivery systems for tissue engineering and regenerative medicine. Using decellularized tissues or purified insoluble collagen as a starting material, the techniques can be applied to synthesize a broad array of tissue-specific bioscaffolds with customizable geometries. The approach involves mechanical processing and mild enzymatic digestion to yield an ECM suspension that is used to fabricate the three-dimensional foams or microcarriers through controlled freezing and lyophilization procedures. These pure ECM-derived scaffolds are highly porous, yet stable without the need for chemical crosslinking agents or other additives that may negatively impact cell function. The scaffold properties can be tuned to some extent by varying factors such as the ECM suspension concentration, mechanical processing methods, or synthesis conditions. In general, the scaffolds are robust and easy to handle, and can be processed as tissues for most standard biological assays, providing a versatile and user-friendly 3D cell culture platform that mimics the native ECM composition. Overall, these straightforward methods for fabricating customized ECM-derived foams and microcarriers may be of interest to both biologists and biomedical engineers as tissue-specific cell-instructive platforms for in vitro and in vivo applications.

The tissue-specific extracellular matrix (ECM) is a key mediator of cell function. This article describes methods for synthesizing pure ECM-derived foams and microcarriers that are stable in culture without the need for chemical crosslinking for applications in advanced 3D in vitro cell culture models or as pro-regenerative bioscaffolds.

细胞外基质衍生的泡沫和微载体如组织特异性细胞培养和交付平台的制造

Cell function is mediated by interactions with the extracellular matrix (ECM), which has complex tissue-specific composition and architecture. The focus ...

Fabrication and Characterization of Griffithsin-modified Fiber Scaffolds for Prevention of Sexually Transmitted Infections

Electrospun fibers (EFs) have been widely used in a variety of therapeutic applications; however, they have only recently been applied as a technology to prevent and treat sexually transmitted infections (STIs). Moreover, many EF technologies focus on encapsulating the active agent, relative to utilizing the surface to impart biofunctionality. Here we describe a method to fabricate and surface-modify poly(lactic-co-glycolic) acid (PLGA) electrospun fibers, with the potent antiviral lectin Griffithsin (GRFT). PLGA is an FDA-approved polymer that has been widely used in drug delivery due to its outstanding chemical and biocompatible properties. GRFT is a natural, potent, and safe lectin that possesses broad activity against numerous viruses including human immunodeficiency virus type 1 (HIV-1). When combined, GRFT-modified fibers have demonstrated potent inactivation of HIV-1 in vitro. This manuscript describes the methods to fabricate and characterize GRFT-modified EFs. First, PLGA is electrospun to create a fiber scaffold. Fibers are subsequently surface-modified with GRFT using 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS)chemistry. Scanning electron microscopy (SEM) was used to assess the size and morphology of surface-modified formulations. Additionally, a gp120 or hemagglutinin (HA)-based ELISA may be used to quantify the amount of GRFT conjugated to, as well as GRFT desorption from the fiber surface. This protocol can be more widely applied to fabricate fibers that are surface-modified with a variety of different proteins.

This manuscript describes the procedure to fabricate and characterize Griffithsin-modified poly(lactic-co-glycolic acid) electrospun fibers that demonstrate potent adhesive and antiviral activity against human immunodeficiency virus type 1 infection in vitro. Methods used to synthesize, surface-modify, and characterize the resulting morphology, conjugation, and desorption of Griffithsin from surface-modified fibers are described.

Griffithsin 修饰纤维支架预防性传播感染的制备与表征

Electrospun fibers (EFs) have been widely used in a variety of therapeutic applications; however, they have only recently been applied as a technology to ...

Agarose-based Tissue Mimicking Optical Phantoms for Diffuse Reflectance Spectroscopy

This protocol describes how to make agarose-based tissue-mimicking phantoms and demonstrates how to determine their optical properties using a conventional optical system with an integrating sphere. Measuring systems for the acquisition of the diffuse reflectance and total transmittance spectra are constructed with a broadband white light source, a light guide, an achromatic lens, an integrating sphere, a sample holder, an optical fiber probe, and a multi-channel spectrometer. An acrylic mold consisting of two rectangular acrylic pieces and a U-shaped acrylic piece is constructed to create an epidermal phantom and a dermal phantom with whole blood. The application of a sodium dithionite (Na2S2O4) solution to the dermal phantom enables the researcher to deoxygenate hemoglobin in red blood cells distributed in the dermal phantom. The inverse Monte Carlo simulation with the diffuse reflectance and total transmittance spectra measured by a spectrometer with an integrating sphere is performed to determine the absorption coefficient spectrum &#181;a(&#955;) and the reduced scattering coefficient spectrum &#181;s&#39;(&#955;) of each layer phantom. A two-layered phantom mimicking the diffuse reflectance of human skin tissue is also demonstrated by piling up the epidermal phantom on the dermal phantom.

Here, we demonstrate how agarose-based tissue-mimicking optical phantoms are made and how their optical properties are determined using a conventional optical system with an integrating sphere.

基于琼脂糖组织模拟光学幽灵的漫反射光谱研究

This protocol describes how to make agarose-based tissue-mimicking phantoms and demonstrates how to determine their optical properties using a ...

Manufacturing Abdominal Aorta Hydrogel Tissue-Mimicking Phantoms for Ultrasound Elastography Validation

Ultrasound (US) elastography, or elasticity imaging, is an adjunct imaging technique that utilizes sequential US images of soft tissues to measure the tissue motion and infer or quantify the underlying biomechanical characteristics. For abdominal aortic aneurysms (AAA), biomechanical properties such as changes in the tissue's elastic modulus and estimates of the tissue stress may be essential for assessing the need for the surgical intervention. Abdominal aortic aneurysms US elastography could be a useful tool to monitor AAA progression and identify changes in biomechanical properties characteristic of high-risk patients.
A preliminary goal in the development of an AAA US elastography technique is the validation of the method using a physically relevant model with known material properties. Here we present a process for the production of AAA tissue-mimicking phantoms with physically relevant geometries and spatially modulated material properties. These tissue phantoms aim to mimic the US properties, material modulus, and geometry of the abdominal aortic aneurysms. Tissue phantoms are made using a polyvinyl alcohol cryogel (PVA-c) and molded using 3D printed parts created using computer aided design (CAD) software. The modulus of the phantoms is controlled by altering the concentration of PVA-c and by changing the number of freeze-thaw cycles used to polymerize the cryogel. The AAA phantoms are connected to a hemodynamic pump, designed to deform the phantoms with the physiologic cyclic pressure and flows. Ultra sound image sequences of the deforming phantoms allowed for the spatial calculation of the pressure normalized strain and the identification of mechanical properties of the vessel wall. Representative results of the pressure normalized strain are presented.

Here we describe a method to manufacture aneurysmal, aortic tissue-mimicking phantoms for the use in testing ultrasound elastography. The combined use of computer-aided design (CAD) and 3-dimensional (3D) printing techniques produce aortic phantoms with predictable, complex geometries to validate the elastographic imaging algorithms with controlled experiments.

超声弹性成像验证腹主动脉水凝胶组织模拟幻影的研制

Ultrasound (US) elastography, or elasticity imaging, is an adjunct imaging technique that utilizes sequential US images of soft tissues to measure the ...