我们感谢 Ms. 索尼娅 Dufentester, Ms. Elke 韦伯和 Mrs. Klaudia Niepagenkämper 为优秀的技术援助。

根据德国动物保护法 (LANUV), 所有动物实验均经 Landesamt Natur、Umwelt 和 Verbraucherschutz (Nordrhein) Westfalen-Tierschutzgesetz。
1. 材料和实验装置
<ol>
	<li>动物护理。
	<ol>
		<li>使用性别和年龄匹配的任何 DSS 敏感株 (例如, C57BL/6) 在20-25 克体重的小鼠。</li>
		<li>根据当地的动物保育指南, 计划每组至少五只小鼠, 并将小鼠置于家中。</li>
		<li>提供标准的鼠粮和蒸压饮用水ad 随意。</li>
		<li>在扫描前至少三天去除标准的 chow, 并将其换成无苜蓿草, 以减少腔荧光。</li>
	</ol>
	</li>
	<li>急性 DSS 诱导的结肠炎。
	<ol>
		<li>溶解2克的 DSS (分子量〜 4万 Da) 在100毫升的蒸压饮用水中获得 2% (瓦特/v 溶液)。</li>
		<li>只用 DSS 解决方案来填满小鼠的饮水供应, 并估计每只老鼠每天5毫升液体。提供相同的饮用水没有 DSS 的控制小鼠10。 注意: 每天监测老鼠直到实验结束。根据当地适用的动物指南, 安乐的老鼠会失去超过20% 的初始体重或变得奄奄一息 (即,持续驼背的姿势, 减少运动, 呼吸吃力, 明显直立的皮毛)福利.</li>
	</ol>
	</li>
	<li>荧光介导的层析成像的制备。
	<ol>
		<li>标签所需的抗体 (例如,大鼠 anti-mouse F4/80) 与荧光染料 (例如, Cyanine7, λ激发: 750 nm, λ发射: 776 nm), 如制造商的协议所述。透析纯化抗体在适当的透析膜 (孔大小和 #60; 50-100 kDa) 反对 1 L 0.15 M 氯化钠为至少 2 h 或隔夜。<ol>
			<li>将抗体转移到0.1 米 NaHCO 的 1 L,3 , 透析至少2小时。</li>
			<li>在二甲基亚砜 (10.8 µL/毫克抗体) 中溶解所需数量的荧光染料, 并将其添加到抗体溶液中。使用荧光染料在20倍的摩尔过剩, 以达到 dye-to-protein 的比例为1:3。</li>
			<li>在黑暗中孵育在4° c 为 1 h. 通过透析对1升0.15 米钠或使用 PD-10 脱盐柱和解决磷酸盐缓冲盐水 (PBS) 中的体内应用去除未标记抗体。</li>
			<li>用分光光度法测定最终抗体浓度和标记比值。</li>
			<li>测量250-330 纳米吸收的蛋白质浓度, 并考虑染料的额外吸收。校正最大吸收在280毫微米 (蛋白质) 由11% 最大吸收在750毫微米 (下步) 为 Cyanine7 标记。</li>
			<li>测量化合物 (通常为 1:10) 在 250-800 nm 的稀释量, 并提取 Cyanine7 在 750 nm 的浓度。</li>
			<li>确定 dye-to-protein 比为: 染料或抗体 = 最大吸收在750毫微米/20万/(最大吸收在280毫微米-0.11 x 最大吸收在750毫微米)/17万。</li>
			<li>将抗体溶液保持在4° c, 并将其从光中屏蔽以避免在注射前漂白。</li>
		</ol>
		</li>
		<li>在注射前立即将所需的抗体溶液容积装入无菌注射器中, 直至使用。</li>
		<li>根据示踪药代动力学确定探针注射的最佳时机和扫描程序。</li>
		<li>麻醉使用 1.5-2.5% 吸入的异氟醚在氧气中, 或将它们安全地放置在一个专门的抑制剂尾静脉注射的标记抗体的小鼠。</li>
		<li>对于全长抗体, 在扫描前注入至少24小时的标记抗体, 如用于小鼠结肠炎的 anti-mouse F4/80 巨噬细胞可视化。通过尾静脉注射小鼠 (静脉注射) 与标记抗体的数量对应的 2.0 nmol 染料。</li>
		<li>使用同样标记的不抗体 (例如,大鼠 IgG 或与主要抗体遗传相对应的另一种同种) 作为同种控制, 剂量相当于特定探针的量。 注: 在注射控制化合物后的体内扫描结果可以作为对特定探针成像数据的解释的参考。</li>
		<li>使用电动剃须刀将动物皮毛剃毛在腹部区域, 以减少对光的反射和吸收。</li>
	</ol>
	</li>
</ol>
2. 技术设备
<ol>
	<li>使用兽医荧光介导的层析成像 (裂变材料) 装置的小动物荧光 (图 4)。</li>
	<li>通过点击 "新研究" 按钮为每个项目创建一个新的研究, 并在研究描述中包括相关的示踪剂, 包括成像参数和剂量供将来参考。</li>
	<li>在本研究中, 通过单击 "新建学习组" 按钮, 根据各自的研究设计 (例如,为特定的跟踪器和不同种控件) 创建研究组。装备每个研究小组与动物的各自数量。</li>
	<li>校准跟踪器构造的系统。<ol>
		<li>对每批示踪剂进行校准, 使标签的变化正常化, 并从 OI 数据中进行定量测量。</li>
		<li>按照仪器制造商的指导, 对各个系统进行校准;在选择 "添加新的跟踪器" 时, 系统将通过步骤提供指南。在探针中提供了应用抗体溶液的稀释和示踪剂的绝对浓度。</li>
		<li>对于裂变材料, 使用一种组织模仿的定义厚度和吸收特性 (类似于重要组织) 的幻像, 并用特定体积的抗体溶液填充。在裂变材料装置上测量这一点。 注: 系统将使用所提供探头的参考测量, 连同给定的浓度, 从未来的活体测量中计算绝对示踪剂浓度。</li>
	</ol>
	</li>
	<li>使用土炕检测盒, 温度为42° c。 注意: 这可以防止小鼠在检查过程中变得体温过低。</li>
</ol>
3. 动物麻醉
<ol>
	<li>使用连续流动的 1.5-2%% 异氟醚 ([2-氯-2-(氟)-11, 1-中和--乙烷]) 和 1.5 L O2/分钟麻醉老鼠。</li></ol>使用专门设计的啮齿动物麻醉吸入系统 (异氟醚蒸发器), 以方便地控制麻醉深度和减少工作人员暴露。
	<li>将鼠标放在防漏感应室中, 并打开用于异氟醚供应的蒸发器 (100% (v/五), 5% 氧, 3 升/分)。监测鼠标, 直到它是横卧和无意识的。</li>
	<li>将它放在检查盒中进行断层扫描, 通过鼻锥连续吸入异氟醚, 剂量为100% 伏/伏, 1.5% 氧, 1.5 升/分钟, 在检查过程中尽量减少运动伪影。对于持续时间超过5分钟的手术, 应将眼膏涂抹于小鼠眼, 以防止角膜损伤。</li>
	<li>通过检查反应来评估麻醉深度。把老鼠放在背上;如果麻醉足够了, 老鼠就不应该转过身来。在脚趾间轻轻地捏老鼠;如果麻醉足够, 腿将不会被撤回 (手术耐受阶段)。</li>

4. 荧光介导的断层扫描
注: 根据需要, 对本研究中使用的裂变材料体系 (见资料表) 中的替代荧光反射成像设备或裂变物质系统, 采用以下具体的细节进行调整。
<ol>
	<li>将麻醉小鼠放回检查盒中。</li>
	<li>执行扫描过程。<ol>
		<li>将卡带插入成像系统并立即关闭, 以确保连续麻醉。从以前创建的研究组中选择适当的示例。从下拉菜单中选择所管理的跟踪器, 以确保正确计算示踪剂浓度值。</li>
		<li>在适当波长 (720 nm Cyanine7) 上获取荧光反射图像, 用于扫描规划, 并通过单击 "获取图像" 按钮来勾勒扫描字段。</li>
		<li>请注意, 扫描字段显示为荧光反射图像上的叠加。将其调整到感兴趣的区域 (例如,冒号或腹部), 避免空气或剩余毛皮的区域。根据图像目标, 通过在右侧的菜单中选择 "粗到中" 和 "精细扫描-字段分辨率", 在扫描字段中设置图像数据点的数目。 注意: 请记住, 一个精细的扫描字段可能提供更好的空间分辨率, 其代价是扫描时间大大延长。</li>
		<li>单击 "扫描", 在选定的波长处启动数据/图像采集。 注: 扫描时间为中细扫描整个腹部将约5分钟;在这段时间内, 每个数据点分别由激励激光照射, 并记录产生的荧光。</li>
		<li>在扫描结束时从成像盒中取出动物, 让动物完全恢复, 然后再将其放回笼子。</li>
		<li>在实验过程中, 如果认为需要在不同的时间点 (例如,天 0, 5, 和 9-10 (实验结束)), 重复的裂变材料, 但考虑体内抗体的积累, 因此增加了背景荧光信号。</li>
	</ol>
	</li>
</ol>
5. 扫描后
<ol>
	<li>将鼠标放在一个单独的笼子里, 放在一盏红光灯下的纸巾上, 监视鼠标是否有不适或苦恼的迹象, 直到完全恢复。当完全清醒时, 将鼠标放回各自的笼子里。</li>
	<li>安乐在实验结束时, 通过将 CO2传递到隔离器, 剂量为 100% (v/v)、100% 和3升/分。不要填充的商会与 co2,作为一个突然暴露在高浓度 co2可能会导致遇险。验证死亡后, 鼠标停止呼吸由随后的第二模式的安乐死, 如快速颈椎脱位。</li>
	<li>通过腹部剖腹术外植体结肠, 并对说明结肠进行活体扫描, 如步骤 4.2.1-4.2.4 所述。在准备进一步的分析之前, 用 Metzenbaum 的外科剪刀纵向打开每个结肠, 用生理盐水彻底冲洗。</li>
	<li>使用手术刀切割从远端结肠0.5 厘米的片段, 并立即放置在1.5 毫升低温管。将其冷冻在液氮中, 贮存在-70 ° c, 直至进一步使用 (例如,过氧化物 (过氧化物酶) 测量)。</li>
	<li>使用木棍和卷起剩余的纵向打开的结肠从远端到近端, 与黏膜向外 ("瑞士卷技术"), 为组织学分析。将准备工作放置在固定剂中 (请参见材料表), 并在-80 ° c14处冻结。</li>
</ol>
6. 数据重建和解释
<ol>
	<li>使用各自成像软件的重建工具, 从原始成像数据中创建3D 荧光分布图;扫描将自动添加到重建工具中, 当扫描时, 会选择 "添加到重建队列" 功能。<ol>
		<li>否则, 从相应的学习和学习组下的下拉菜单中选择扫描, 右键单击扫描, 然后选择 "添加到重建"。</li>
	</ol>
	</li>
	<li>要进一步分析, 请将数据集加载到分析软件中。从顶栏的下拉菜单中选择相应的研究, 然后从右侧的菜单中选择学习组和单个动物;所有对此动物进行的扫描将显示。选择正确的扫描, 然后单击 "加载"。 注: 示踪剂分布的3D 重建将在左侧出现, 作为最初获得的荧光反射图像的叠加。该模型可以旋转和放大, 以便于分析。</li>
	<li>在重建的3D 地图上确定不标签堆积 (例如,肝或尿囊) 的病灶, 并与靶组织 (如肠或肠道) 区分。</li>
	<li>从顶部栏中选择最适合目标的 ROI 形状。通过在分析软件中放置相应的测量工具, 将目标组织标记为感兴趣的区域 (ROI);该软件将提供一个荧光强度的 ROI, 以及 (微微) 摩尔量的示踪剂, 扫描已被校准。</li>
	<li>在适当的 roi 中选择跟踪器的总数量, 将其归为 roi 大小, 作为与炎症浸润 (组织学) 相关的疾病活动评估的最合适的等价物。 注: ROI 的其他功能可以在适当时作为特定模型的代表选择。</li>
</ol>
7。Ex 活体分析
<ol>
	<li>苏木精和红素染色和免疫荧光染色。
	<ol>
		<li>Deparaffinize 部分通过放置在70% 乙醇 (乙醇) 为2分钟;之后用蒸馏水冲洗。染色用苏木精溶液5分钟, 随后用温水冲洗10分钟。</li>
		<li>用蒸馏水冲洗, counterstain 用曙红溶液2分钟, 再用蒸馏水冲洗。</li>
		<li>放置在70% 乙醇, 96% 乙醇, 99% 乙醇, 和二甲苯 (2 次) 为2分钟, 以脱水和清除部分。装载树脂安装介质。</li>
	</ol>
	</li>
	<li>免疫荧光染色。
	<ol>
		<li>使用先前获得的组织切片 ("瑞士卷") 进行免疫荧光染色, 并准备7µm 的冷冻切片。</li>
		<li>在5.0% 只大鼠血清中阻断丙酮固定和冰冻结肠切片10分钟, 并在夜间用稀释的 (1/500 伏/v) 化原 anti-mouse F4/80 抗体进行孵育。在三缓冲盐水 (TBS) 三次洗涤部分和孵育与亲和-FITC (1:100 v/v) 在 PBS 或 BSA (0.1% 瓦特/v) 隔夜在4° c。</li>
		<li>用 4, 6-diamidino-2-吲 (DAPI, 1:1000 伏/v) 来洗涤 TBS 和染色的部分, 以达到对比。在共焦显微镜下分析荧光图像 (见材料表; 40x 放大倍率; 过滤器立方体 N2.1 与励磁过滤器 515-560) 和计数 F4/80-positive 细胞每个大功率领域。 注意: 考虑使用相同的克隆和格式的抗体结合在体内的裂变材料和验尸组织学分析, 如免疫组织化学或免疫荧光染色, 根据预期的目标。为检测 F4/80-positive 巨噬细胞在体内和ex 体内, 使用了不同的格式。</li>
	</ol>
	</li>
	<li>髓过氧化物酶在结肠样品中的测量。
	<ol>
		<li>使用新鲜获得的, PBS 冲洗的结肠样品或解冻的样品进行过氧化物酶测量。在 ELISA 试剂盒中提供的溶解缓冲液中, 称量所有样品和质组织 (见材料表), 每毫克组织的裂解缓冲液量为20µL。</li>
		<li>几种为十五年代 (超声频率:20 赫, 力量:70 W) 和离心样品为10分钟在 200 x g 和4° c。</li>
		<li>根据制造商的描述, 使用商业上可用的 ELISA 试剂盒 (参见材料表)。执行重复的测试。</li>
	</ol>
	</li>
</ol>

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作者没有什么可透露的。

尽管近年来医学成像技术发展迅速, 但在早期的发育阶段, 我们在检测炎症过程或肿瘤以及其他疾病方面的能力仍然有限。然而, 这对于了解肿瘤的生长, 侵袭, 或转移的发展和细胞过程中的炎症性疾病和退化, 心血管和免疫疾病的发展至关重要。虽然传统的成像技术依赖于物理或生理参数, 分子成像使特定分子标记的可视化在体内20。
对于小动物的成像,...

动物模型被广泛应用于科学研究, 许多非侵入性的程序都存在于监测疾病的活动和生命力, 如体重变化的量化或血液、尿液和粪便的分析。但是, 这些只是间接的代理参数, 也受个体的可变性。它们必须经常通过对组织标本的验尸分析来补充, 防止在重复的时间点进行连续观察, 并直接观察生理或病理过程在体内。成熟的小动物成像技术已经出现, 包括横断面成像, 光学成像和内窥镜, 这使这些过程的直接可视化, 也允许重复分析相同的动物1,2,3. 此外, 在同一动物中反复监测各种疾病的可能性可能会减少所需的动物数量, 从动物伦理学的角度来看, 这可能是可取的。
几种不同的光学成像技术存在于体内荧光成像。最初, 共聚焦成像被用来研究表面和地下荧光事件4,5。然而, 最近, 允许进行定量的三维组织评估的层析系统已经开发成6。这是通过开发荧光探针, 在近红外线 (近红外光谱) 频谱发出光, 提供低吸收, 灵敏的探测器, 单色光源7。虽然传统的切削成像技术, 如计算机断层扫描 (CT), 磁共振成像 (MRI), 或超声 (美国), 主要依赖于物理参数和可视化形态学, 光学成像可以提供额外的信息使用内生或外源荧光探针的基础分子过程8。
分子生物学的进步有助于为越来越多的目标提供智能和有针对性的荧光分子探针的生成。例如, 受体介导的摄取和分布在给定的目标区域可以可视化使用 carbocyanine 衍生物标记抗体9。大量可用的抗体, 可被标记为功能作为特定的示踪剂在其他无法进入的身体区域, 提供了前所未有的洞察力分子和细胞的过程中的模型的肿瘤和神经退行性,心血管、免疫学和炎症性疾病7。
在这项研究中, 我们描述了使用荧光介导的层析成像在小鼠结肠炎模型。葡聚糖硫酸钠 (DSS) 诱导结肠炎是一种标准的化学诱导的小鼠肠道炎症模型, 类似炎症性肠病 (IBD)10。这是特别有用的评估先天免疫系统对发展的肠道炎症的贡献11。由于单核和巨噬细胞的招募、活化和浸润是 IBD 发病机制的关键步骤, 因此它们的招募和渗透动力学的可视化是监测的关键, 例如, 影响潜在的治疗性药物在临床前设置12。我们描述了诱导的 DSS 结肠炎, 并演示了利用荧光分子层析成像技术对巨噬细胞浸润肠黏膜的特性, 为单核/巨噬细胞标记 F4/80 的具体可视化13. 此外, 我们还说明了辅助和补充程序, 如抗体标签;实验装置;对所获得的图像进行分析和解释, 与常规读数相关, 如疾病活动指数、流式细胞仪和组织学分析、免疫组织化学。我们讨论这项技术的局限性和与其他成像方式的比较。

<table><tbody><tr><td>&lt;strong&gt;Reagents&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Alfalfa-free diet</td><td>Harlan Laboritories, Madison, USA</td><td>2014</td><td></td></tr><tr><td>Bepanthen eye ointment</td><td>Bayer, Leverkusen, Germany</td><td>80469764</td><td></td></tr><tr><td>Dextran sulphate sodium (DSS)</td><td>TdB Consulatancy, Uppsala, Sweden</td><td>DB001</td><td></td></tr><tr><td>Eosin</td><td>Sigma - Aldrich, Deisenhofen, Germany</td><td>E 4382</td><td></td></tr><tr><td>Ethylenediaminetetraacetic acid (EDTA)&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Sigma - Aldrich, Deisenhofen, Germany</td><td>E 9884</td><td></td></tr><tr><td>Florene 100V/V</td><td>Abbott, Wiesbaden, Germany</td><td>B506</td><td></td></tr><tr><td>Haematoxylin&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Sigma - Aldrich, Deisenhofen, Germany</td><td>HHS32-1L</td><td></td></tr><tr><td>O.C.T. Tissue Tek compound&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Sakura, Zoeterwonde, Netherlands</td><td>4583</td><td>fixative for histological analyses</td></tr><tr><td>Phosphate buffered saline, PBS</td><td>Lonza, Verviers, Belgium</td><td>4629</td><td></td></tr><tr><td>Sodium Chloride 0,9%</td><td>Braun, Melsungen, Germany</td><td>5/12211095/0411</td><td></td></tr><tr><td>Sodium bicarbonate powder</td><td>Sigma Aldrich Deisenhofen, Germany</td><td>S5761</td><td></td></tr><tr><td>Standard diet</td><td>Altromin, Lage, Germany</td><td>1320</td><td></td></tr><tr><td>Tissue-Tek Cryomold</td><td>Sakura, Leiden, Netherlands</td><td>4566</td><td></td></tr><tr><td>Hemoccult (guaiac paper test)</td><td>Beckmann Coulter, Germany</td><td>3060</td><td></td></tr><tr><td>Biotin rat-anti-mouse anti-F4/80 antibody</td><td>Serotec, Oxford, UK</td><td>MCA497B</td><td></td></tr><tr><td>Biotin rat-anti-mouse anti-GR-1&amp;nbsp;</td><td>BD Pharmingen, Heidelberg Germany</td><td>553125</td><td></td></tr><tr><td>Streptavidin-Alexa546</td><td>Molecular Probes, Darmstadt, Germany</td><td>S-11225</td><td>excitation/emission maximum:&amp;nbsp; 556/573nm</td></tr><tr><td>Anti-CD11b rat-anti-mouse antibody TC</td><td>Calteg, Burlingame, USA</td><td>R2b06</td><td></td></tr><tr><td>Purified anti-mouse F4/80 antibody</td><td>BioLegend, London, UK</td><td>123102</td><td></td></tr><tr><td>DAPI</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>D9542</td><td></td></tr><tr><td>FITC-conjugated anti-Ly6C rat-anti-mouse antibody</td><td>BD Pharmingen, Heidelberg, Germany</td><td>553104</td><td></td></tr><tr><td>FACS buffer</td><td>BD Pharmingen, Heidelberg, Germany</td><td>342003</td><td></td></tr><tr><td>Cy7 NHS Ester</td><td>GE Healthcare&amp;nbsp;Europe, Freiburg, Germany</td><td>PA17104</td><td></td></tr><tr><td>MPO ELISA</td><td>Immundiagnostik AG, Bensheim, Germany</td><td>K 6631B</td><td></td></tr><tr><td>Cy5.5 labeled anti-mouse F4/80 antibody</td><td>BioLegend, London, UK</td><td>123127</td><td>ready to use labelled Antibodies (alternative)</td></tr><tr><td>Anti-Mouse F4/80 Antigen PerCP-Cyanine5.5</td><td>eBioscience, Waltham, USA</td><td>45-4801-80</td><td>ready to use labelled Antibodies (alternative)</td></tr><tr><td>DMSO (Dimethyl sulfoxide)</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>67-68-5</td><td></td></tr><tr><td>Isoflurane</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>792632</td><td></td></tr><tr><td>Ethanol</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>64-17-5</td><td></td></tr><tr><td>Bovine Serum Albumins (BSA)</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>A4612</td><td></td></tr><tr><td>Tris Buffered Saline Solution (TBS)</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>SRE0032</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>FACS Calibur Flow Cytometry System</td><td>BD Biosciences GmbH, Heidelberg, Germany</td><td></td><td></td></tr><tr><td>FMT 2000 In Vivo Imaging System</td><td>PerkinElmer Inc., Waltham, MA, USA</td><td>FMT2000</td><td></td></tr><tr><td>True Quant 3.1 Imaging Analysis Software</td><td>PerkinElmer Inc., Waltham, MA, USA</td><td>included in FMT2000</td><td></td></tr><tr><td>Leica DMLB Fluorescent Microscope</td><td>Leica,&amp;nbsp; 35578 Wetzlar, Germany&amp;nbsp;</td><td>DMLB</td><td></td></tr><tr><td>Bandelin Sonopuls HD 2070</td><td>Bandelin, 12207 Berlin, Germany</td><td>HD 2070</td><td>ultrasonic homogenizer</td></tr><tr><td>Disposable scalpel No 10</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>Z692395-10EA</td><td></td></tr><tr><td>Metzenbaum scissors 14cm</td><td>Ehrhardt Medizinprodukte GmbH, Geislingen, Germany</td><td>22398330</td><td></td></tr><tr><td>luer lock syringe 5ml</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>Z248010</td><td></td></tr><tr><td>syringe needles</td><td>Sigma-Aldrich, Deisenhoffen, Germany</td><td>Z192368&amp;nbsp;</td><td></td></tr><tr><td>Falcon Tube 50ml</td><td>BD Biosciences, Erembodegem, Belgium</td><td>352070</td><td></td></tr></tbody></table>

fluorescence-mediated tomography for the detection and quantification of macrophage-related murine intestinal inflammation

小鼠疾病模型对科学研究是必不可少的。然而, 许多诊断工具, 如内窥镜或断层成像, 在动物模型中并不经常使用。传统的实验读数通常依赖于宰后和前体分析, 从而防止个体的随访检查, 增加所需的研究动物数量。荧光介导的断层扫描使非侵入性的, 重复的, 定量的, 三维荧光探针的评估。它是高度敏感的, 并允许使用分子制造商, 这使得特定的检测和表征明确的分子目标。特别是, 有针对性的探针代表了一个创新的工具, 分析基因活化和蛋白表达在炎症, 自身免疫疾病, 感染, 血管疾病, 细胞迁移, 肿瘤,等。在这篇文章中, 我们提供了 step-by 的步骤说明, 对这种复杂的成像技术的在体内检测和定性的炎症 (即, F4/80-positive 巨噬细胞浸润) 在广泛使用的小鼠模型肠道炎症。该技术也可用于其他研究领域, 如免疫细胞或干细胞追踪。

结肠炎的评估:
DSS 诱导的结肠炎是一种化学诱导的小鼠肠道炎症模型, 类似于人 IBD, 导致体重下降, 直肠出血, 表面溃疡, 和粘膜损害的敏感的老鼠15。研究先天免疫系统对肠道炎症发育的贡献特别有用10,11。为了能诱发 colitic 炎症, 小鼠在整个实验过程中...

Watch this Scientific Journal Video about Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation at JoVE.com

Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

目标特定的探针代表了一种用于分析分子机制的创新工具, 例如各种疾病的蛋白质表达 (例如,炎症、感染和肿瘤发生)。在这项研究中, 我们描述了一个定量的三维层析成像评估肠道巨噬细胞浸润的小鼠结肠炎模型使用 F4/80-specific 荧光介导的断层扫描。

荧光介导的断层扫描对巨噬细胞相关小鼠肠道炎症的检测和定量

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not ...

fluorescence-mediated-tomography-for-detection-quantification

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

8.1K 次观看.  University Hospital Münster. 目标特定的探针代表了一种用于分析分子机制的创新工具, 例如各种疾病的蛋白质表达 (例如,炎症、感染和肿瘤发生)。在这项研究中, 我们描述了一个定量的三维层析成像评估肠道巨噬细胞浸润的小鼠结肠炎模型使用 F4/80-specific 荧光介导的断层扫描。

视频: Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

Computed Tomography-guided Time-domain Diffuse Fluorescence Tomography in Small Animals for Localization of Cancer Biomarkers

Small animal fluorescence molecular imaging (FMI) can be a powerful tool for preclinical drug discovery and development studies1. However, light absorption by tissue chromophores (e.g., hemoglobin, water, lipids, melanin) typically limits optical signal propagation through thicknesses larger than a few millimeters2. Compared to other visible wavelengths, tissue absorption for red and near-infrared (near-IR) light absorption dramatically decreases and non-elastic scattering becomes the dominant light-tissue interaction mechanism. The relatively recent development of fluorescent agents that absorb and emit light in the near-IR range (600-1000 nm), has driven the development of imaging systems and light propagation models that can achieve whole body three-dimensional imaging in small animals3.
Despite great strides in this area, the ill-posed nature of diffuse fluorescence tomography remains a significant problem for the stability, contrast recovery and spatial resolution of image reconstruction techniques and the optimal approach to FMI in small animals has yet to be agreed on. The majority of research groups have invested in charge-coupled device (CCD)-based systems that provide abundant tissue-sampling but suboptimal sensitivity4-9, while our group and a few others10-13 have pursued systems based on very high sensitivity detectors, that at this time allow dense tissue sampling to be achieved only at the cost of low imaging throughput. Here we demonstrate the methodology for applying single-photon detection technology in a fluorescence tomography system to localize a cancerous brain lesion in a mouse model.
The fluorescence tomography (FT) system employed single photon counting using photomultiplier tubes (PMT) and information-rich time-domain light detection in a non-contact conformation11. This provides a simultaneous collection of transmitted excitation and emission light, and includes automatic fluorescence excitation exposure control14, laser referencing, and co-registration with a small animal computed tomography (microCT) system15. A nude mouse model was used for imaging. The animal was inoculated orthotopically with a human glioma cell line (U251) in the left cerebral hemisphere and imaged 2 weeks later. The tumor was made to fluoresce by injecting a fluorescent tracer, IRDye 800CW-EGF (LI-COR Biosciences, Lincoln, NE) targeted to epidermal growth factor receptor, a cell membrane protein known to be overexpressed in the U251 tumor line and many other cancers18. A second, untargeted fluorescent tracer, Alexa Fluor 647 (Life Technologies, Grand Island, NY) was also injected to account for non-receptor mediated effects on the uptake of the targeted tracers to provide a means of quantifying tracer binding and receptor availability/density27. A CT-guided, time-domain algorithm was used to reconstruct the location of both fluorescent tracers (i.e., the location of the tumor) in the mouse brain and their ability to localize the tumor was verified by contrast-enhanced magnetic resonance imaging.
Though demonstrated for fluorescence imaging in a glioma mouse model, the methodology presented in this video can be extended to different tumor models in various small animal models potentially up to the size of a rat17.

Diffuse fluorescence tomography offers a relatively low-cost and potentially high-throughout approach to preclinical in vivo tumor imaging. The methodology of optical data collection, calibration, and image reconstruction is presented for a computed tomography-guided non-contact time-domain system using fluorescent targeting of the tumor biomarker epidermal growth factor receptor in a mouse glioma model.

电脑断层导引的时域漫荧光体层摄影术在小动物的肿瘤生物标志物的本土化

Small animal fluorescence molecular imaging (FMI) can be a powerful tool for preclinical drug discovery and development studies1. However, light ...

科研

医学

In vivo Imaging Method to Distinguish Acute and Chronic Inflammation

Inflammation is a fundamental aspect of many human diseases. In this video report, we demonstrate non-invasive bioluminescence imaging techniques that distinguish acute and chronic inflammation in mouse models. With tissue damage or pathogen invasion, neutrophils are the first line of defense, playing a major role in mediating the acute inflammatory response. As the inflammatory reaction progresses, circulating monocytes gradually migrate into the site of injury and differentiate into mature macrophages, which mediate chronic inflammation and promote tissue repair by removing tissue debris and producing anti-inflammatory cytokines. Intraperitoneal injection of luminol (5-amino-2,3-dihydro-1,4-phthalazinedione, sodium salt) enables detection of acute inflammation largely mediated by tissue-infiltrating neutrophils. Luminol specifically reacts with the superoxide generated within the phagosomes of neutrophils since bioluminescence results from a myeloperoxidase (MPO) mediated reaction. Lucigenin (bis-N-methylacridinium nitrate) also reacts with superoxide in order to generate bioluminescence. However, lucigenin bioluminescence is independent of MPO and it solely relies on phagocyte NADPH oxidase (Phox) in macrophages during chronic inflammation. Together, luminol and lucigenin allow non-invasive visualization and longitudinal assessment of different phagocyte populations across both acute and chronic inflammatory phases. Given the important role of inflammation in a variety of human diseases, we believe this non-invasive imaging method can help investigate the differential roles of neutrophils and macrophages in a variety of pathological conditions.

In vivo Imaging Method to Distinguish Acute and Chronic Inflammation

We describe a non-invasive imaging method for distinguishing inflammatory stages. Systemic delivery of luminol reveals areas of acute inflammation dependent upon MPO activity in neutrophils. In contrast, injection of lucigenin allows for visualization of chronic inflammation dependent upon Phox activity in macrophages.

在体内成像方法区分急性和慢性炎症

Inflammation is a fundamental aspect of many human diseases. In this video report, we demonstrate non-invasive bioluminescence imaging techniques that ...

免疫与感染

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

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 number of imaging techniques currently available and the progress in instrumentation, there is still a need for highly sensitive probes that are suitable for in vivo imaging. One typical problem of available preclinical fluorescent probes is their rapid clearance in vivo, which reduces their imaging sensitivity. To circumvent rapid clearance, increase number of dye molecules at the target site, and thereby reduce background autofluorescence, encapsulation of the near-infrared fluorescent dye, DY-676-COOH in liposomes and verification of its potential for in vivo imaging of inflammation was done. DY-676 is known for its ability to self-quench at high concentrations. We first determined the concentration suitable for self-quenching, and then encapsulated this quenching concentration into the aqueous interior of PEGylated liposomes. To substantiate the quenching and activation potential of the liposomes we use a harsh freezing method which leads to damage of liposomal membranes without affecting the encapsulated dye. The liposomes characterized by a high level of fluorescence quenching were termed Lip-Q. We show by experiments with different cell lines that uptake of Lip-Q is predominantly by phagocytosis which in turn enabled the characterization of its potential as a tool for in vivo imaging of inflammation in mice models. Furthermore, we use a zymosan-induced edema model in mice to substantiate the potential of Lip-Q in optical imaging of inflammation in vivo. Considering possible uptake due to inflammation-induced enhanced permeability and retention (EPR) effect, an always-on liposome formulation with low, non-quenched concentration of DY-676-COOH (termed Lip-dQ) and the free DY-676-COOH were compared with Lip-Q in animal trials.

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

The use of fluorophores for in vivo imaging can be greatly limited by opsonization, rapid clearance, low detection sensitivity and cytotoxic effects on the host. Encapsulation of fluorophores in liposomes by film hydration and extrusion leads to fluorescence quenching and protection which enables in vivo imaging with high detection sensitivity.

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 ...

生物工程

Hybrid &#181;CT-FMT imaging and image analysis

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to assess the biodistribution of novel probes and to assess disease progression using established molecular probes or reporter genes. The combination with an anatomical modality, e.g., micro computed tomography (&#181;CT), is beneficial for image analysis and for fluorescence reconstruction. We describe a protocol for multimodal &#181;CT-FMT imaging including the image processing steps necessary to extract quantitative measurements. After preparing the mice and performing the imaging, the multimodal data sets are registered. Subsequently, an improved fluorescence reconstruction is performed, which takes into account the shape of the mouse. For quantitative analysis, organ segmentations are generated based on the anatomical data using our interactive segmentation tool. Finally, the biodistribution curves are generated using a batch-processing feature. We show the applicability of the method by assessing the biodistribution of a well-known probe that binds to bones and joints.

Hybrid  &#181;CT-FMT imaging and image analysis

We describe a protocol for hybrid imaging, combining fluorescence-mediated tomography (FMT) with micro computed tomography (&#181;CT). After fusion and reconstruction, we perform interactive organ segmentation to extract quantitative measurements of the fluorescence distribution.

混合μCT-FMT成像和图像分析

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to ...

Real-time Imaging of Myeloid Cells Dynamics in ApcMin/+ Intestinal Tumors by Spinning Disk Confocal Microscopy

Myeloid cells are the most abundant immune cells within tumors and have been shown to promote tumor progression. Modern intravital imaging techniques enable the observation of live cellular behavior inside the organ but can be challenging in some types of cancer due to organ and tumor accessibility such as intestine. Direct observation of intestinal tumors has not been previously reported. A surgical procedure described here allows direct observation of myeloid cell dynamics within the intestinal tumors in live mice by using transgenic fluorescent reporter mice and injectable tracers or antibodies. For this purpose, a four-color, multi-region, micro-lensed spinning disk confocal microscope that allows long-term continuous imaging with rapid image acquisition has been used. ApcMin/+ mice that develop multiple adenomas in the small intestine are crossed with c-fms-EGFP mice to visualize myeloid cells and with ACTB-ECFP mice to visualize intestinal epithelial cells of the crypts. Procedures for labeling different tumor components, such as blood vessels and neutrophils, and the procedure for positioning the tumor for imaging through the serosal surface are also described. Time-lapse movies compiled from several hours of imaging allow the analysis of myeloid cell behavior in situ in the intestinal microenvironment.

Real-time Imaging of Myeloid Cells Dynamics in ApcMin/+ Intestinal Tumors by Spinning Disk Confocal Microscopy

By using transgenic reporter mice and injectable fluorescent labels, long-term intravital spinning disk confocal microscopy enables direct visualization of myeloid cell behavior into intestinal adenoma in the ApcMin/+ colorectal cancer model.

在髓系细胞动态实时成像<em&gt;装甲运兵车<sup&gt;最小/ +</sup</em&gt;肠道肿瘤通过旋转盘共聚焦显微镜

Myeloid cells are the most abundant immune cells within tumors and have been shown to promote tumor progression. Modern intravital imaging techniques ...

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions preclinically. However, valid assessment of pathological alterations often requires histological analysis, and when performed ex vivo, necessitates death of the animal. Therefore in conventional experimental settings, intra-individual follow-up examinations are rarely possible. Thus, development of murine endoscopy in live mice enables investigators for the first time to both directly visualize the gastrointestinal mucosa and also repeat the procedure to monitor for alterations. Numerous applications for in vivo murine endoscopy exist, including studying intestinal inflammation or wound healing, obtaining mucosal biopsies repeatedly, and to locally administer diagnostic or therapeutic agents using miniature injection catheters. Most recently, molecular imaging has extended diagnostic imaging modalities allowing specific detection of distinct target molecules using specific photoprobes. In conclusion, murine endoscopy has emerged as a novel cutting-edge technology for diagnostic experimental in vivo imaging and may significantly impact on preclinical research in various fields.

Murine Endoscopy for In Vivo Multimodal Imaging of Carcinogenesis and Assessment of Intestinal Wound Healing and Inflammation

Small animal imaging techniques allow serial diagnostic examinations and therapeutic interventions in vivo. Recently, the scope of applications has significantly widened and currently includes assessment of colonic tumor development, wound healing and monitoring of inflammation. This protocol illustrates these diverse potential applications of murine endoscopy.

鼠内镜<em&gt;体内</em&gt;发生和肠道的评价多模式成像伤口愈合和炎症

Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

通过轻型显微镜对小鼠急性心肌炎模型中白细胞浸润的定量可视化

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Intravital Imaging of Intraepithelial Lymphocytes in Murine Small Intestine

Intraepithelial lymphocytes expressing &#947;&#948; T cell receptor (&#947;&#948; IEL) play a key role in immune surveillance of the intestinal epithelium. Due in part to the lack of a definitive ligand for the &#947;&#948; T cell receptor, our understanding of the regulation of &#947;&#948; IEL activation and their function in vivo remains limited. This necessitates the development of alternative strategies to interrogate signaling pathways involved in regulating &#947;&#948; IEL function and the responsiveness of these cells to the local microenvironment. Although &#947;&#948; IELs are widely understood to limit pathogen translocation, the use of intravital imaging has been critical to understanding the spatiotemporal dynamics of IEL/epithelial interactions at steady-state and in response to invasive pathogens. Herein, we present a protocol for visualizing IEL migratory behavior in the small intestinal mucosa of a GFP &#947;&#948; T cell reporter mouse using inverted spinning disk confocal laser microscopy. Although the maximum imaging depth of this approach is limited relative to the use of two-photon laser-scanning microscopy, spinning disk confocal laser microscopy provides the advantage of high speed image acquisition with reduced photobleaching and photodamage. Using 4D image analysis software, T cell surveillance behavior and their interactions with neighboring cells can be analyzed following experimental manipulation to provide additional insight into IEL activation and function within the intestinal mucosa.

We describe a method to visualize GFP-labeled &#947;&#948; IELs using intravital imaging of murine small intestine by inverted spinning disk confocal microscopy. This technique enables the tracking of live cells within the mucosa for up to 4 h and can be used to investigate a variety of intestinal immune-epithelial interactions. &#160;

毛里小肠内皮淋巴细胞的内质成像

Intraepithelial lymphocytes expressing &#947;&#948; T cell receptor (&#947;&#948; IEL) play a key role in immune surveillance of the intestinal ...

Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ Hybridization, Lectin Staining, and Imaging

Measuring the localization of microbes within their in vivo context is an essential step in revealing the functional relationships between the microbiota and the vertebrate gut. The spatial landscape of the gut microbiota is tightly controlled by physical features - intestinal mucus, crypts, and folds - and is affected by host-controlled properties such as pH, oxygen availability, and immune factors. These properties limit the ability of commensal microbes and pathogens alike to colonize the gut stably. At the micron-scale, microbial organization determines the close-range interactions between different microbes as well as the interactions between microbes and their host. These interactions then affect large-scale organ function and host health.
This protocol enables the visualization of the gut microbiota spatial organization from distances between cells to organ-wide scales. The method is based on fixing gut tissues while preserving intestinal structure and mucus properties. The fixed samples are then embedded, sectioned, and stained to highlight specific bacterial species through fluorescence in situ hybridization (FISH). Host features, such as mucus and host cell components, are labeled with fluorescently labeled lectins. Finally, the stained sections are imaged using a confocal microscope utilizing tile-scan imaging at high magnification to bridge the micron to centimeter length scales. This type of imaging can be applied to intestinal sections from animal models and biopsies from human tissues to determine the biogeography of the microbiota in the gut in health and disease.

Visualization of Gut Microbiota-host Interactions via Fluorescence In Situ Hybridization, Lectin Staining, and Imaging

This streamlined protocol details a workflow to detect and image bacteria in complex tissue samples, from fixing the tissue to staining microbes with fluorescent in situ hybridization.

通过荧光 原位 杂交、凝集素染色和成像可视化肠道微生物群与宿主的相互作用

Measuring the localization of microbes within their in vivo context is an essential step in revealing the functional relationships between the microbiota ...

通过荧光报告基因定量进行全器官病原体追踪

High&#45;Resolution Three&#45;Dimensional Whole&#45;Organ Tomography of Microbial Infections

Most infections take place within three-dimensional host tissues with intricate anatomy and locally varying host physiology. The positioning of pathogen cells within this diverse environment significantly affects their stress levels, responses, fate, and contribution to the overall progression of the disease and treatment failure. However, due to the technical difficulties in locating &#181;m-sized pathogen cells within cm-sized host organs, this area of research has been relatively unexplored. Here, we present a method for addressing this challenge. We employ serial two-photon tomography and AI-enhanced image analysis to locate individual Salmonella cells throughout the entire spleen, liver lobes, and whole lymph nodes of infected mice. Using fluorescent reporters and in vivo antibody administration, the replication rate of single Salmonella cells, their local interaction with specific immune cells, and bacterial responses to antibiotics can be determined. These methodologies open avenues for a comprehensive examination of infections, their prevention, and treatment within the three-dimensional tissue context.

作者聚焦:揭示细菌对组织中抗生素和免疫系统的反应

Here, we describe a procedure that enables organ-wide detection of pathogenic bacteria during infection and quantification of fluorescent reporter activities.

微生物感染的高分辨率三维全器官断层扫描

Most infections take place within three-dimensional host tissues with intricate anatomy and locally varying host physiology. The positioning of pathogen ...

荧光介导的断层扫描对巨噬细胞相关小鼠肠道炎症的检测和定量

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此视频中的章节

电脑断层导引的时域漫荧光体层摄影术在小动物的肿瘤生物标志物的本土化

在体内成像方法区分急性和慢性炎症

荧光淬一个脂质体包封的近红外荧光团的作为一个工具

混合μCT-FMT成像和图像分析

在髓系细胞动态实时成像

鼠内镜

通过轻型显微镜对小鼠急性心肌炎模型中白细胞浸润的定量可视化

毛里小肠内皮淋巴细胞的内质成像

通过荧光原位杂交、凝集素染色和成像可视化肠道微生物群与宿主的相互作用

微生物感染的高分辨率三维全器官断层扫描

荧光介导的断层扫描对巨噬细胞相关小鼠肠道炎症的检测和定量

摘要

探索更多视频

此视频中的章节

电脑断层导引的时域漫荧光体层摄影术在小动物的肿瘤生物标志物的本土化

在体内成像方法区分急性和慢性炎症

荧光淬一个脂质体包封的近红外荧光团的作为一个工具

混合μCT-FMT成像和图像分析

在髓系细胞动态实时成像

鼠内镜

通过轻型显微镜对小鼠急性心肌炎模型中白细胞浸润的定量可视化

毛里小肠内皮淋巴细胞的内质成像

通过荧光 原位 杂交、凝集素染色和成像可视化肠道微生物群与宿主的相互作用

微生物感染的高分辨率三维全器官断层扫描

通过荧光原位杂交、凝集素染色和成像可视化肠道微生物群与宿主的相互作用