Name: a bright nir-ii fluorescence probe for vascular and tumor imaging
Uploaded: 2023-03-17T14:55:00
Description: Watch this Scientific Journal Video about A Bright NIR-II Fluorescence Probe for Vascular and Tumor Imaging at JoVE.com

这项工作得到了国家自然科学基金（82273796，82111530209）、中央指导地方科技发展专项资金（XZ202202YD0021C、XZ202102YD0033C、XZ202001YD0028C）、湖北省科技创新重点项目（2020BAB058）、中央高校基础研究经费和西藏自治区COVID-19科技发展防控项目的资助。

用于NIR-II成像研究的动物实验在武汉大学动物实验中心进行，该中心已获得国际实验动物护理协会（AALAC）的称号。所有动物研究均按照《中国动物福利委员会实验动物护理使用指南》进行，并经武汉大学动物实验中心动物护理与使用委员会（IACUC）批准。
本研究使用6周龄的雌性BALB / c裸鼠（~20g）。
1. 近红外II成像制备
<ol><li>将市售的黑色?...

<ol>
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NIR-I荧光成像在一定程度上可用于肿瘤和血管成像，但由于NIR-I荧光团的最大发射波长（<900nm）有限，导致组织渗透性和肿瘤信号背景比为33，34。成像分辨率差和低可能会导致成像反馈治疗的结果与实际治疗效果之间的偏差。此外，大多数NIR-I荧光团的光学稳定性差，代谢极快，导致成像过程不稳定。由于NIR-I荧光团的组织渗透率低且不稳定性，在肿瘤?...

荧光成像是基础研究中常用的分子成像工具，也常用于指导临床肿瘤切除手术1。荧光成像的基本原理是利用相机接收样品（组织、器官等）照射后激光发出的荧光。2. 该过程在几毫秒内完成3.荧光成像波长可分为紫外（200-400 nm）、可见光区（400-700 nm）、近红外 I（NIR-I，700-900 nm）和近红外 II（NIR-II，1000-1700 nm）4，5，6。由于生物组织中的血红蛋白、黑色素、脱氧血红蛋白、胆红素等内源分子对可见光区域的光有较强的吸收和散射作用，因此光的穿透力和灵敏度大大降低，对可见光波长的荧光成像产生不利影响7，8，9。
NIR-II荧光成像具有低光子吸收和散射，高成像速度和高图像对比度（或灵敏度）10，11。随着荧光波长的增加，荧光在生物组织中的吸收和散射逐渐减小，NIR-II区的自发荧光极低12。因此，NIR-II窗口显着增加了组织的穿透深度，并获得更高的分辨率和信噪比13，14，15。NIR-II窗口可进一步细分为NIR-IIa（1300-1400 nm）和NIR-lIb（1500-1700 nm）窗口16。迄今为止，已有多个里程碑式的NIR-II材料报道，包括无机材料单壁碳纳米管、稀土纳米颗粒、量子点，以及有机材料半导体聚合物纳米颗粒、小分子染料、聚集诱导发光材料等。1，17，18，19，20，21，22.无机纳米材料易在肝脏、脾脏等体内蓄积，具有潜在的长期生物毒性23.有机小分子荧光团具有代谢快、毒性低、易修饰、结构清晰等优点，是临床上最有前景的探针24。
NIR-II光学成像系统也是荧光生物成像的关键组成部分，因为它可以有效地从NIR-II探头收集NIR-II荧光信号，从而提供精确的功能，解剖和分子图像25，26。NIR-II成像系统主要包括短波红外相机、长通（LP）滤光片、激光器和计算机处理器。 体内NIR-II荧光成像被认为是阐明疾病机制和生命本质的最可行的成像方法之一27，28，29。NIR-II成像技术已广泛应用于癌细胞检测、动态成像、体内靶向示踪、靶向治疗等生物医学领域，尤其在肿瘤学研究方面30、31。然而，考虑到NIR-II.成像技术对成像探头和仪器的高技术要求，也给不同领域的研究人员的实际应用带来了困惑和限制。因此，本文详细介绍了NIR-II成像探头的制备和NIR-II成像的应用。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Anhydrous pyridine</td>
			<td>Perimed&#160;</td>
			<td>110-86-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Anhydrous sodium sulfate</td>
			<td>China national medicines Co.,Ltd</td>
			<td>SY006376</td>
			<td></td>
		</tr>
		<tr>
			<td>Black cardboard</td>
			<td>Suzhou Yingrui Optical Technology Co., Ltd</td>
			<td>AO00158</td>
			<td></td>
		</tr>
		<tr>
			<td>Column chromatography</td>
			<td>Energy Chemical</td>
			<td>E080498</td>
			<td></td>
		</tr>
		<tr>
			<td>Diphenylphosphine palladium dichloride</td>
			<td>Sigma-Aldrich</td>
			<td>B2161-1g</td>
			<td></td>
		</tr>
		<tr>
			<td>DSPE-PEG2000</td>
			<td>Ponsure</td>
			<td>PS-E1</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s modified eagle medium&#160;</td>
			<td>Gibco</td>
			<td>8121587</td>
			<td></td>
		</tr>
		<tr>
			<td>EGTA</td>
			<td>Biofroxx</td>
			<td>EZ6789D115</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Gibco</td>
			<td>2166090RP</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>GLPBIO</td>
			<td>GC45487-1</td>
			<td></td>
		</tr>
		<tr>
			<td>K2CO3</td>
			<td>Macklin</td>
			<td>P816305-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>N. N &#39;- dimethylformamide</td>
			<td>China national medicines Co.,Ltd</td>
			<td>02-12-1968</td>
			<td></td>
		</tr>
		<tr>
			<td>NIR-II imaging instrument</td>
			<td>Suzhou Yingrui Optical Technology Co., Ltd</td>
			<td>16011109</td>
			<td></td>
		</tr>
		<tr>
			<td>N-sulfenanilide</td>
			<td>Enerry chemical&#160;</td>
			<td>1250030-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>PdCl2(dppf)2CH2Cl2</td>
			<td>TCI&#160;</td>
			<td>B2064-1g</td>
			<td></td>
		</tr>
		<tr>
			<td>penicillin-streptomycin</td>
			<td>Gibco</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetrahydrofuran</td>
			<td>China national medicines Co.,Ltd</td>
			<td>M005197</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetratriphenylphosphine palladium</td>
			<td>Immochem</td>
			<td>1021232-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetratriphenylphosphine palladium</td>
			<td>Sigma-Aldrich</td>
			<td>1021232-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>Tributyltin chloride</td>
			<td>Immochem</td>
			<td>QH004335</td>
			<td></td>
		</tr>
		<tr>
			<td>Trimethylchlorosilane</td>
			<td>China national medicines Co.,Ltd</td>
			<td>40060560</td>
			<td></td>
		</tr>
	</tbody>
</table>

a bright nir-ii fluorescence probe for vascular and tumor imaging

近红外II（NIR-II，1000-1700 nm）荧光成像作为一种新兴的成像技术，因其高灵敏度、深层组织穿透性和具有时空分辨率的卓越成像，在生物医学领域具有巨大的潜力。然而，促进NIR-II荧光成像在医学、药学等一些急需领域实施的方法，一直困扰着相关研究人员。该协议详细描述了具有D-A-D（供体-受体-供体）骨架的NIR-II荧光分子探针HLY1的构建和生物成像应用。HLY1具有良好的光学性能和生物相容性。此外，使用NIR-II光学成像设备对小鼠进行NIR-II血管和肿瘤成像。采集实时高分辨率NIR-II荧光图像，指导肿瘤和血管疾病的检测。从探针制备到数据采集，成像质量大大提高，保证了活体成像中用于数据记录的NIR-II分子探针的真实性。

通过NIR-II成像仪器测定水悬浮HLY1点的荧光强度和亮度。90% f w THF/H2O 混合物中 HLY1 的荧光强度是 THF 溶液中的五倍，这表明 HLY1 具有突出的 AIE 特征（图 1B）。此外，HLY1点在1，500 nm LP滤光片下发出强烈的荧光信号，表明HLY1点可用于NIR-IIb成像（图1D）。HLY1点的最大吸收和最大发射波长分别为740 nm和1，040 nm（图2A</str...

Watch this Scientific Journal Video about A Bright NIR-II Fluorescence Probe for Vascular and Tumor Imaging at JoVE.com

A Bright NIR-II Fluorescence Probe for Vascular and Tumor Imaging

本协议描述了使用NIR-II光学成像装置对小鼠进行详细的实时NIR-II荧光成像操作。

用于血管和肿瘤成像的明亮NIR-II荧光探针

As an emerging imaging technology, near-infrared II (NIR-II, 1000-1700 nm) fluorescence imaging has significant potential in the biomedical field, owing ...

通过近红外到荧光纳米探针实现了高分辨率的血管和肿瘤成像，为指导血管疾病和癌症提供了一种准确有效的方法。NIR-II窗口中的活体成像使我们能够深入研究活体，为生物医学研究和我们的临床应用创造了机会。通过在成像设备的载体中心放置市售的黑色纸板来准备近红外II或NIR-II成像。然后，将样品放在黑色纸板的顶部，使其位于载体的中心。选择合适的过滤器后，执行平台高度调整。长按托架控制台控制区域触摸屏界面上的平台向上选项以将其向上移动。并长按平台向下移动。合成NIR-II染料HLY1后，以DSPE-PEG-2K为包封基质，通过纳米沉淀法制备HLY1点。为此，将装有 10 毫克 DSPE-PEG-2K 溶液的烧杯放入 9 毫升水中，在 25 摄氏度下进行超声处理。然后，将一毫克HLY1溶解在一毫升四氢呋喃或THF中。并将溶液缓慢加入到进行超声处理的DSPE-PEG-2K水溶液中。随后，通过透析从混合物中去除THF。将上述溶液以7，100G的超滤离心浓缩10分钟后，将其置于4摄氏度的冰箱中以备将来使用。对于体内成像，将HLY1点静脉注射到麻醉小鼠中。三分钟后，继续使用光学NIR-II成像系统对小鼠全身的血管进行NIR-II荧光成像。进一步关注小鼠的头部以收集脑血管成像。将光学NIR-II成像系统的仪器参数设置为90毫瓦/平方厘米和808纳米激光。在小鼠中注射HLY1点五分钟后收集图像，并使用ImageJ软件处理数据。含有90%水的90%THF溶液中HLY1的荧光强度是THF溶液的五倍，表明HLY1具有明显的聚集诱导发射或AIE特征。HLY1点在1， 500纳米低通或LP滤波器下发出强烈的荧光信号，证明其适用于NIR-IIb成像。HLY1点的最大吸收和发射波长分别为740纳米和1， 040纳米。此外，通过动态光散射确定HLY1点的流体动力学尺寸为145纳米。在用HLY1点给药的小鼠中，在1， 500纳米LP滤光片下通过NIR-II成像清楚地识别后肢的脑血管和微血管。此外，荷瘤小鼠的四个T1肿瘤也通过NIR-II成像清晰可见，表明HLY1点的通透性和保留性或EPR效应增强。所有这些结果表明，HLY1点是一种明亮的NIR-II荧光探针，适用于血管和肿瘤成像。在体内和NIR-II荧光成像下制备水悬浮纳米探针是该过程最重要的部分。

Research

JoVE Journal

Medicine

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

体内近红外荧光（NIRF）血管内分子成像炎性斑块，多式联运方针成像动脉粥样硬化

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

科研

医学

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

生物发光和微型CT成像监测肿瘤转移和溶骨性病变

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

多光子成像在口腔鳞状细胞癌原位小鼠模型的肿瘤细胞的侵袭

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

在肿瘤血管生成中使用荧光共聚焦电视显微镜活体成像

Fibered confocal fluorescence in vivo imaging with a fiber optic bundle uses the same principle as fluorescent confocal microscopy. It can excite ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

一种新型的高分辨率<em&gt;在体内</em&gt;成像技术研究肿瘤生长及治疗颅内结构的动态响应

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

药物输送到微型计算机断层扫描成像缺血后肢血管灌注与微滤渗透

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Computerized Dynamic Posturography for Postural Control Assessment in Patients with Intermittent Claudication

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic ...

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

在皮下肿瘤模型的评估肿瘤浸润白细胞亚群

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Establishment of a Human Multiple Myeloma Xenograft Model in the Chicken to Study Tumor Growth, Invasion and Angiogenesis

建立一个人类多发性骨髓瘤异种移植模型中的鸡，研究肿瘤的生长，侵袭和血管生成

Multiple myeloma (MM), a malignant plasma cell disease, remains incurable and novel drugs are required to improve the prognosis of patients. Due to the ...

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

之后，正常小鼠70％部分肝切除血管和实质再生的可视化

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...