Name: terahertz imaging and characterization protocol for freshly excised breast cancer tumors
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这项工作由国家卫生研究院（NIH）奖（R15CA208798）资助，部分由美国国家科学基金会（NSF）奖1408007资助。脉冲THz系统的资金是通过NSF/MRI奖1228958获得的。我们承认使用国家疾病研究交流会 （NDRI） 在 NIH 赠款 U42OD11158 的支持下采购的纸巾。我们还感谢与俄克拉荷马州立大学俄克拉荷马动物疾病诊断实验室合作，对本工作处理的所有组织进行组织病理学程序。

该协议遵循阿肯色大学环境健康和安全部门制定的所有要求。
1. 设置组织处理区域
<ol><li>取一个不锈钢金属托盘，用生物危害袋盖住它，如图1所示。任何生物组织处理都将在托盘区域（即组织处理区域）内进行。</li>
	<li>在托盘周围准备实验室钳、纸巾、纸巾、滤纸包、组织染料瓶、漂白瓶和乙醇瓶，以便在需要时方便携带。在生物危害物质表面保留...

<ol>
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H., 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=Terahertz+spectroscopy+for+the+assessment+of+burn+injuries+in+vivo.">Terahertz spectroscopy for the assessment of burn injuries in vivo.</a> Journal of Biomedical Optics. 18 (7), 077004 (2013).</li><li>Sy, S., 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=Terahertz+spectroscopy+of+liver+cirrhosis:+investigating+the+origin+of+contrast.">Terahertz spectroscopy of liver cirrhosis: investigating the origin of contrast.</a> Physics in Medicine and Biology. 55 (24), 7587-7596 (2010).</li><li>Yu, C., Fan, S., Sun, Y., Pickwell-Macpherson, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+potential+of+terahertz+imaging+for+cancer+diagnosis:+A+review+of+investigations+to+date.">The potential of terahertz imaging for cancer diagnosis: A review of investigations to date.</a> Quantitative Imaging in Medicine and Surgery. 2 (1), 33-45 (2012).</li><li>El-Shenawee, M., Vohra, N., Bowman, T., Bailey, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+detection+in+excised+breast+tumors+using+terahertz+imaging+and+spectroscopy.">Cancer detection in excised breast tumors using terahertz imaging and spectroscopy.</a> Biomedical Spectroscopy and Imaging. 8 (1-2), 1-9 (2019).</li><li>Yamaguchi, S., 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=Brain+tumor+imaging+of+rat+fresh+tissue+using+terahertz+spectroscopy.">Brain tumor imaging of rat fresh tissue using terahertz spectroscopy.</a> Scientific Reports. 6 (30124), 1-6 (2016).</li><li>Rong, 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=Terahertz+in-line+digital+holography+of+human+hepatocellular+carcinoma+tissue.">Terahertz in-line digital holography of human hepatocellular carcinoma tissue.</a> Scientific Reports. 5 (8445), 1-6 (2015).</li><li>Park, J. Y., Choi, H. J., Nam, G., Cho, K., Son, J. <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+Dual-Modality+Terahertz+/+Magnetic+Resonance+Imaging+Using+Superparamagnetic+Iron+Oxide+Nanoparticles+as+a+Dual+Contrast+Agent.">In Vivo Dual-Modality Terahertz / Magnetic Resonance Imaging Using Superparamagnetic Iron Oxide Nanoparticles as a Dual Contrast Agent.</a> IEEE Transactions on Terahertz Science and Technology. 2 (1), 93-98 (2012).</li><li>Ji, Y. B., 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=Feasibility+of+terahertz+reflectometry+for+discrimination+of+human+early+gastric+cancers.">Feasibility of terahertz reflectometry for discrimination of human early gastric cancers.</a> Biomedical Optics Express. 6 (4), 1413-1421 (2015).</li><li>Bowman, T., 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+Phantom+Study+of+Terahertz+Spectroscopy+and+Imaging+of+Micro-+and+Nano-diamonds+and+Nano-onions+as+Contrast+Agents+for+Breast+Cancer.">A Phantom Study of Terahertz Spectroscopy and Imaging of Micro- and Nano-diamonds and Nano-onions as Contrast Agents for Breast Cancer.</a> Biomedical Physics and Engineering Express. 3 (5), 055001 (2017).</li><li>Chavez, T., Bowman, T., Wu, J., Bailey, K., El-Shenawee, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+Terahertz+Imaging+for+Excised+Breast+Cancer+Tumors+with+Image+Morphing.">Assessment of Terahertz Imaging for Excised Breast Cancer Tumors with Image Morphing.</a> Journal of Infrared, Millimeter, and Terahertz Waves. 39 (12), 1283-1302 (2018).</li><li>Bowman, T. C., El-Shenawee, M., Campbell, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Terahertz+Imaging+of+Excised+Breast+Tumor+Tissue+on+Paraffin+Sections.">Terahertz Imaging of Excised Breast Tumor Tissue on Paraffin Sections.</a> IEEE Transactions on Antennas and Propagation. 63 (5), 2088-2097 (2015).</li><li>Bowman, T., El-Shenawee, M., Campbell, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Terahertz+transmission+vs+reflection+imaging+and+model-based+characterization+for+excised+breast+carcinomas.">Terahertz transmission vs reflection imaging and model-based characterization for excised breast carcinomas.</a> Biomedical Optics Express. 7 (9), 3756-3783 (2016).</li><li>Bowman, T., Wu, Y., Gauch, J., Campbell, L. K., El-Shenawee, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Terahertz+Imaging+of+Three-Dimensional+Dehydrated+Breast+Cancer+Tumors.">Terahertz Imaging of Three-Dimensional Dehydrated Breast Cancer Tumors.</a> Journal of Infrared, Millimeter, and Terahertz Waves. 38 (6), 766-786 (2017).</li><li>Bowman, T., 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=Pulsed+terahertz+imaging+of+breast+cancer+in+freshly+excised+murine+tumors.">Pulsed terahertz imaging of breast cancer in freshly excised murine tumors.</a> Journal of Biomedical Optics. 23 (2), 026004 (2018).</li><li>Bowman, T., Vohra, N., Bailey, K., El-Shenawee, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Terahertz+tomographic+imaging+of+freshly+excised+human+breast+tissues.">Terahertz tomographic imaging of freshly excised human breast tissues.</a> Journal of Medical Imaging. 6 (2), 023501 (2019).</li><li>Vohra, N., 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=Pulsed+Terahertz+Reflection+Imaging+of+Tumors+in+a+Spontaneous+Model+of+Breast+Cancer.">Pulsed Terahertz Reflection Imaging of Tumors in a Spontaneous Model of Breast Cancer.</a> Biomedical Physics and Engineering Express. 4 (6), 065025 (2018).</li><li>Jacobs, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Positive+margins:+the+challenge+continues+for+breast+surgeons.">Positive margins: the challenge continues for breast surgeons.</a> Annals of Surgical Oncology. 15 (5), 1271-1272 (2008).</li><li>Moran, M. S., 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=Society+of+Surgical+Oncology--American+Society+for+Radiation+Oncology+Consensus+Guideline+on+Margins+for+Breast-Conserving+Surgery+With+Whole-Breast+Irradiation+in+Stages+I+and+II+Invasive+Breast+Cancer.">Society of Surgical Oncology--American Society for Radiation Oncology Consensus Guideline on Margins for Breast-Conserving Surgery With Whole-Breast Irradiation in Stages I and II Invasive Breast Cancer.</a> International Journal of Radiation Oncology. 88 (3), 553-564 (2014).</li><li>Fitzgerald, A. 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=Terahertz+Pulsed+Imaging+of+human+breast+tumors.">Terahertz Pulsed Imaging of human breast tumors.</a> Radiology. 239 (2), 533-540 (2006).</li><li>Ashworth, P. C., 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=Terahertz+pulsed+spectroscopy+of+freshly+excised+human+breast+cancer.">Terahertz pulsed spectroscopy of freshly excised human breast cancer.</a> Optics Express. 17 (15), 12444-12454 (2009).</li><li>Doradla, P., Alavi, K., Joseph, C., Giles, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+colon+cancer+by+continuous-wave+terahertz+polarization+imaging+technique.">Detection of colon cancer by continuous-wave terahertz polarization imaging technique.</a> Journal of Biomedical Optics. 18 (9), 090504 (2013).</li><li>Reid, C. B., 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=Terahertz+pulsed+imaging+of+freshly+excised+human+colonic+tissues.">Terahertz pulsed imaging of freshly excised human colonic tissues.</a> Physics in Medicine and Biology. 56 (1), 4333-4353 (2011).</li><li>. Teraview.com Available from: <a target="_blank" href="https://teraview.com">https://teraview.com</a> (2019)</li><li>Orosco, R. K., 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=Positive+Surgical+Margins+in+the+10+Most+Common+Solid+Cancers.">Positive Surgical Margins in the 10 Most Common Solid Cancers.</a> Scientific Reports. 8 (1), 1-9 (2018).</li><li>Bowman, T., 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=Statistical+signal+processing+for+quantitative+assessment+of+pulsed+terahertz+imaging+of+human+breast+tumors.">Statistical signal processing for quantitative assessment of pulsed terahertz imaging of human breast tumors.</a> 2017 42nd International Conference on Infrared, Millimeter, and Terahertz Waves (IRMMW-THz). , 1-2 (2017).</li><li>Gavdush, A. 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=Terahertz+spectroscopy+of+gelatin-embedded+human+brain+gliomas+of+different+grades:+a+road+toward+intraoperative+THz+diagnosis.">Terahertz spectroscopy of gelatin-embedded human brain gliomas of different grades: a road toward intraoperative THz diagnosis.</a> Journal of Biomedical Optics. 24 (2), 027001 (2019).</li></ol>

新鲜组织的有效THz反射成像主要取决于两个关键方面：1） 正确考虑组织处理（第 2 节和第 4.15 节）;和 2） 舞台设置（主要是第 4.11 节）。由于 DMEM 和其他液体的高反射，组织干燥不足会导致反射增加，无法可视化区域。同时，与成像窗口的不良组织接触会在 THz 反射图像中产生环或低反射点，从而掩盖结果。应作出额外努力，确保组织与成像窗口有良好的接触，包括重新定位组织以获得更好的?...

在过去的十年里，Terahertz （THz） 成像和光谱学一直是一个快速增长的研究领域。在 0.1-4 THz 范围内持续开发更高效、更一致的 THz 发射器，使得其应用显著增长1。THz 已显示出希望和显著增长的一个领域是生物医学领域2。THz辐射已被证明是非电离和生物安全在功率水平通常用于分析固定组织3。因此，THz成像和光谱学被用来分类和区分各种组织特征，如含水量，以指示烧伤损伤和愈合4，肝硬化5，和癌症在切除组织6,6，7。癌症评估特别涵盖广泛的潜在的临床和外科应用，并已调查脑8，肝脏9，卵巢10，胃肠道11，和乳房7，12，13，14，15，16，17，18，1912,13,14,15,16,17,18的19癌症。7
THz乳腺癌的应用主要侧重于通过保证金评估支持乳房保护手术或乳房切除术。乳房切除术的目的是切除肿瘤和周围健康组织的一小层，与切除整个乳房的全乳房切除术形成鲜明对比。切除组织的手术边缘通过病理学进行评估，一旦样品固定在正式内，切片，嵌入石蜡，并安装在显微镜幻灯片上的4 μm×5μm切片。这个过程可能非常耗时，如果观察到正差值20，则需要稍后进行二次外科手术。美国放射肿瘤学会目前的指导方针将这种正差定义为癌细胞接触表面边缘墨水21。高吸收水合组织的THz成像主要限于表面成像，根据组织类型进行一些不同的穿透，足以满足快速边缘评估的手术需求。快速分析手术设置期间的保证金条件将大大降低手术费用和后续手术率。迄今为止，THz已被证明在区分癌症和健康组织在正式固定，石蜡嵌入（FFPE）组织的有效，但需要额外的调查，以提供可靠的癌症检测在新鲜切除的组织7。
该协议详细介绍了对从生物库获得的新切除人体组织样本进行THz成像和光谱的步骤。建立在新鲜切除的人类乳腺癌组织的THz应用很少用于发表的研究77，18，22，23，18,22,23特别是由研究组不与医院整合。使用新切除组织在其他癌症应用中同样罕见，大多数非乳腺癌的例子都报道了结肠癌24，25。24,原因之一是FFPE组织块比新切除的组织更容易获得和处理，除非用于研究的THz系统是手术工作流程的一部分。同样，大多数商业实验室THz系统不准备处理新鲜组织，而那些确实还处于使用细胞系生长阶段或刚刚开始查看动物模型中切除的组织。要将 THz 应用于术中设置，需要提前为新鲜组织开发成像和表征步骤，以便分析不会干扰执行标准病理学的能力。对于并非本质上意味着术中应用的应用，新鲜组织的表征仍然是一个具有挑战性的步骤，必须加以解决，以在体内应用和分化方面发挥作用。
这项工作的目的是为使用商业 THz 系统的新切除组织 THz 应用提供指南。该协议是在THz成像和光谱系统26为鼠乳腺癌肿瘤13，17，19，17,并13扩展到人类手术组织从生物组197,7，18。虽然该协议是为乳腺癌生成的，但相同的概念可以应用于类似的THz成像系统和其他类型的固体肿瘤癌症，这些癌症在手术中得到治疗，而手术的成功取决于保证金评估27。由于新切除组织上已公布的 THz 结果相当少，这是作者所知的第一部专注于 THz 成像和表征新鲜组织处理协议的工作。

<table>
	<tbody>
		<tr>
			<td>70% isopropyl alcohol</td>
			<td>VWR</td>
			<td>89108-162</td>
			<td>Contains 70% USP grade isopropanol and 30% USP grade deionized water</td>
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		<tr>
			<td>Alconox powder detergent</td>
			<td>VWR</td>
			<td>21835-032</td>
			<td>Concentrated detergent to remove organic contaminants from glass, metal, stainless steel, porcelain, ceramic, plastic, rubber, and fiberglass</td>
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		<tr>
			<td>Bio Hazard Bags</td>
			<td>Fisher Scientific</td>
			<td>19-033-712</td>
			<td>Justrite FM-Approved Biohazard Waste Container Replacement Bags</td>
		</tr>
		<tr>
			<td>Cardboard holder</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Scrap cardboard to keep tissue imaging face intact when immersed in formalin</td>
		</tr>
		<tr>
			<td>Centrifuge Tubes</td>
			<td>VWR</td>
			<td>10026-078</td>
			<td>Centrifuge Tubes with Flat Caps, Conical-Bottom, Polypropylene, Sterile, Standard Line</td>
		</tr>
		<tr>
			<td>Cotton Swabs</td>
			<td>Walmart</td>
			<td>551398298</td>
			<td>Q-tips Original Cotton Swabs used to dye the tissue</td>
		</tr>
		<tr>
			<td>Ethyl Alcohol</td>
			<td>VWR</td>
			<td>71002-426</td>
			<td>KOPTECH Pure (undenatured) anhydrous (200 proof/100%) ethyl alcohol</td>
		</tr>
		<tr>
			<td>Eye protection goggles</td>
			<td>VWR</td>
			<td>89130-918</td>
			<td>Kimberly-clark professional safety glasses</td>
		</tr>
		<tr>
			<td>Face Mask</td>
			<td>VWR</td>
			<td>95041-774</td>
			<td>DUKAL Corporation surgical masks</td>
		</tr>
		<tr>
			<td>Filter paper</td>
			<td>Sigma Aldrich</td>
			<td>Z240087</td>
			<td>Whatman grade 1 cellulose filters</td>
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		<tr>
			<td>Formalin solution</td>
			<td>Sigma Aldrich</td>
			<td>HT501128-4L</td>
			<td>10% neutral buffered formalin</td>
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		<tr>
			<td>Human freshly excised tumors (Infilterating Ductal Carcinoma (IDC))</td>
			<td>National Disease Research Interchange (NDRI biobank</td>
			<td>N/A</td>
			<td>A protocol is signed with the NDRI for the type of tumors required</td>
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			<td>IRADECON Bleach solution</td>
			<td>VWR</td>
			<td>89234-816</td>
			<td>Pre-diluted Sodium Hypochlorite Bleach solution</td>
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			<td>KIMTECH SCIENCE wipes</td>
			<td>VWR</td>
			<td>21905-026</td>
			<td>Kimberly-clark professional Kim wipes</td>
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			<td>Laboratory Coat</td>
			<td>VWR</td>
			<td>10141-342</td>
			<td>This catalog number is for medium size coat</td>
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			<td>Laboratory tweezers/Forceps</td>
			<td>VWR</td>
			<td>82027-388</td>
			<td>Any laboratory tweezers can be used as long as it does not damage the tissue</td>
		</tr>
		<tr>
			<td>Liquid sample holder (two quartz windows with a 0.1 mm teflon spacer)</td>
			<td>TeraView, Ltd</td>
			<td>N/A</td>
			<td>1&#34; diameter, and 0.1452&#34; thick quartz windows</td>
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		<tr>
			<td>Nitrile hand gloves</td>
			<td>VWR</td>
			<td>82026-426</td>
			<td>This catalog number is for medium size gloves</td>
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			<td>Nitrogen cylinder</td>
			<td>Airgas</td>
			<td>NI UHP300</td>
			<td>NITROGEN UHP GR 5.0 SIZE 300</td>
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			<td>Paper towel</td>
			<td>VWR</td>
			<td>14222-321</td>
			<td>11 x 8.78&#34; Sheets, 1 Ply</td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>VWR</td>
			<td>52858-076</td>
			<td>Flexible thermoplastic. Rolled, waterproof sheet interwound with paper to prevent self-adhesion.</td>
		</tr>
		<tr>
			<td>Petri Dish</td>
			<td>VWR</td>
			<td>470210-568</td>
			<td>VWR Petri Dish, Slippable, Mono Plate (undivided bottom)</td>
		</tr>
		<tr>
			<td>Polystyrene Plate</td>
			<td>Home Depot</td>
			<td>1S11143A</td>
			<td>~ 10 x 10 cm square piece cut from a 11&#34; x 14&#34; x 0.05&#34; Non-glare styrene sheet</td>
		</tr>
		<tr>
			<td>ScanAcquire Software</td>
			<td>TeraView, Ltd</td>
			<td>N/A</td>
			<td>System Software for THz reflection imaging measurements</td>
		</tr>
		<tr>
			<td>Stainless steel low-profile blade (#4689)</td>
			<td>VWR</td>
			<td>25608-964</td>
			<td>Tissue-Tek Accu-Edge Disposable Microtome Blades</td>
		</tr>
		<tr>
			<td>Stainless steel metal tray</td>
			<td>Quick Medical</td>
			<td>10F</td>
			<td>Polar Ware Stainless Steel Medical Instrument Trays</td>
		</tr>
		<tr>
			<td>Tissue Marking Dyes</td>
			<td>Ted Pella, Inc</td>
			<td>Yellow Dye #27213-1 
			Red Dye #27213-2 
			Blue Dye #27213-4</td>
			<td>Used to orient excised tissue samples 
			sent to the histopathology laboratory</td>
		</tr>
		<tr>
			<td>TPS Spectra 3000</td>
			<td>TeraView, Ltd</td>
			<td>N/A</td>
			<td>THz imaging and spectroscopy system</td>
		</tr>
		<tr>
			<td>TPS Spectra Software</td>
			<td>TeraView, Ltd</td>
			<td>N/A</td>
			<td>System Software for THz transmission spectroscopy measurements</td>
		</tr>
	</tbody>
</table>

terahertz imaging and characterization protocol for freshly excised breast cancer tumors

本手稿提供了一个协议，使用脉冲太赫兹成像和光谱技术处理、描述和成像新切除的人类乳腺肿瘤。该协议涉及在正常入射处的太赫兹传输模式和斜角为30°的太赫兹反射模式。收集的实验数据表示电场的时域脉冲。通过切除组织上的固定点传输的太赫兹电场信号通过分析模型进行处理，以提取组织的折射率和吸收系数。利用步进电机扫描仪，从肿瘤的每个像素反射出太赫兹发射脉冲，提供不同组织区域的平面图像。图像可以在时间或频域中显示。此外，利用每个像素的折射率和吸收系数的提取数据，提供肿瘤的图谱太赫兹图像。该协议表明癌组织和健康组织之间的明显区别。另一方面，由于肿瘤表面存在气泡和液体，不遵守协议可能会导致图像发出嘈杂或不准确。该协议提供了一种乳腺肿瘤的手术边缘评估方法。

图9中介绍了根据上述人类乳腺癌肿瘤标本#ND14139从生物库接收的THz成像结果。根据病理报告，#ND14139肿瘤是I/II级渗透导管癌（IDC），通过左乳房结块切除术手术从一名49岁的妇女那里获得。肿瘤的照片如图9A所示，图9B中的病理图像，以及协议中使用方程（1）获得的THz功率光谱图像，如图9C?...

Watch this Scientific Journal Video about Terahertz Imaging and Characterization Protocol for Freshly Excised Breast Cancer Tumors at JoVE.com

Terahertz Imaging and Characterization Protocol for Freshly Excised Breast Cancer Tumors

新鲜切除的人类乳腺癌肿瘤的特点是使用太赫兹光谱和成像遵循新鲜的组织处理协议。组织定位被考虑在内，以实现有效的表征，同时及时为将来的术中应用提供分析。

新切除乳腺癌肿瘤的Terahertz成像和表征协议

This manuscript presents a protocol to handle, characterize, and image freshly excised human breast tumors using pulsed terahertz imaging and spectroscopy ...

该协议使用 Terahertz 成像和光谱学来评估乳腺肿瘤的手术边缘，旨在大幅减少第二次手术需求。Terahertz 成像可用于根据其光学特性，以非电离和生物安全的方式区分癌组织和非癌组织。这项技术是为乳腺癌边缘评估开发的，但它适用于需要手术切除和边缘评估的其他实体肿瘤。此方法允许对FFPE组织块内部进行成像，而无需物理切片，便于在任何深度精确识别肿瘤边缘。该协议要求执行组织处理和阶段平衡等关键步骤，如果没有视觉演示，这些步骤可能难以理解。在处理纸巾覆盖不锈钢金属托盘之前，使用生物危害袋，将所有适当的实验室材料放在覆盖托盘的方便范围内。接下来，将新鲜肿瘤样本转移到托盘上的培养皿中，并使用毛检查来选择肿瘤的一侧，该侧足够平坦，血液很少，血管少。然后，将要成像的一侧放在一级滤纸上，以去除任何多余的需求介质，并清除液体或分泌物的组织，在纸张饱和时将肿瘤重新定位到干燥点。在保持肿瘤干燥五分钟时，将一个 1.2 毫米厚的聚苯乙烯板放在直径约 37 毫米的扫描窗上，然后将扫描窗口和聚苯乙烯板放在 Terahertz 成像仪的样品阶段。在主窗口中单击固定点扫描图标以激活 Terahertz 天线，并开始从聚苯乙烯板上的单个点发送和接收反射的 Terahertz 信号。单击电机舞台对话框图标。电机控制窗口将打开。要将主窗口中聚苯乙烯的反射脉冲居中，请单击光学延迟轴箭头以调整光学延迟轴。单击"数据采集设置"按钮打开"数据采集设置对话"窗口，将光学延迟值从 5 伏更改为 4 伏。使用微米刻度调整扫描台的垂直位置，直到次脉冲的最小值最强，并调整电机控制窗口中轴的光延迟，将主反射置于所测量的反射信号范围之外。要调平样品级并记录参考信号，请单击电机控制对话按钮以打开电机控制窗口，重新定位电机控制和主软件窗口，使时域信号在调整电机位置时可见。要使用电机控制窗口中的以下步骤对 A 轴进行升级，请将 x 轴的值从零更改为负 10 并点击 Enter。舞台将移动到 A 轴的负 10 毫米位置，信号位置将观察到主窗口的变化。使用可调微米刻度将信号的最小峰值移回上一个垂直位置，并更改 A 轴值为正 10。单击"输入"。舞台将移动到 A 轴上的加号 10 毫米位置，信号将再次观察到移动。请注意，信号从上一个位置移动的方向和距离，将 A 轴值更改回负 10。信号将返回到原始垂直位置。在扫描阶段的 A 轴上旋转水平螺钉，将信号移向同一方向的距离加倍，该阶段从原始位置移动。使用扫描阶段的千分尺将信号移回原始位置，然后重复调整，直到正 10 和减 10 时的信号在峰值中相等，两个位置的峰值都聚焦在原始位置。实现 A 轴的电平后，将 A 轴值更改为零，然后对 B 轴重复相同的过程。两个轴都平平后，将 A 轴和 B 轴返回为零毫米，并包围电机控制窗口，然后验证信号是否处于其原始位置，以防信号发生一些变化。若要在"数据获取属性"窗口中将此信号记录为参考，请将平均值更改为 5，然后将所有其他参数保留为默认设置。然后单击新引用。平均计数器将从零计数到 20。计数器达到 20 后，将平均值更改为 1，然后单击"确定"。来自聚苯乙烯的反射信号将保存为任何后续扫描的参考。设置所有参数后，将成像窗口从扫描阶段转移到组织处理区，然后将肿瘤安装到聚苯乙烯板上。用钳子去除肿瘤内的任何气泡，或轻轻地将样品卷到聚苯乙烯上，直到将气隙降至最低。定期在测试样品周围放置吸收间隔器，并在肿瘤上方放置另一个聚苯乙烯板。轻轻按压肿瘤表面尽可能平，并胶带下肿瘤和聚苯乙烯板排列在样品窗口。扁平的组织表面和与聚苯乙烯板良好的接触至关重要，因为气泡、多余的液体和不平整的表面会破坏肿瘤组织成像。翻转样品窗口并拍摄肿瘤的照片，以记录其方向。将示例窗口返回到扫描阶段，然后单击图像参数对话按钮以打开图像采集参数窗口。设置轴一个最小值、轴一最大值、轴 2 个最小值和轴数两最大值，以完全包围肿瘤在成像窗口中的位置。为成像扫描设置轴一步轴和两步轴到 0.2 毫米。然后，在"测量"菜单下，选择"回飞 2D 扫描"，并创建一个文件夹和文件名，在其中将扫描数据保存在弹出窗口中。在这项分析中，评估了从一名49岁妇女通过左乳房切除术获得的一至二级渗透导管癌。将太赫兹图像与病理学图像关联后，很明显，癌症区域表现出比脂肪区域更高的反射。断层成像显示，随着频率的增加，脂肪像素中癌症的计算吸收系数值增加，癌症像素显示的值高于两个频率的脂肪值。相比之下，两个组织之间的折射率随着频率的增加而降低。同一肿瘤的透射光谱分析揭示了从肿瘤中提取的两个部分提取的吸收系数和折射率的频率范围。请注意，对组织处理不当会导致误导性的成像结果，例如，由于未处理肿瘤样本中存在过量液体，导致肿瘤中存在更大的癌症。可以在 Terahertz 图像数据上执行其他表征，以获取有关组织频率相关属性的信息，并可执行统计分析以进行自动图像分割。该技术已导致成像和光谱算法的发展，用于研究导管癌的原位特征，以区分低级肿瘤和致命的高等级肿瘤。请记住，要谨慎处理癌组织和形式溶液，并且任何与肿瘤组织接触的系统元素或工具必须适当清洁或丢弃。

Research

JoVE Journal

Engineering

Terahertz Microfluidic Sensing Using a Parallel-plate Waveguide Sensor

太赫兹微流控测量中的一个平行板波导传感器

Refractive index (RI) sensing is a powerful noninvasive and label-free sensing technique for the identification, detection and monitoring of microfluidic ...

科研

工程学

Design, Fabrication, and Experimental Characterization of Plasmonic Photoconductive Terahertz Emitters

电浆光电导太赫兹发射器的设计，制造和实验表征

In this video article we present a detailed demonstration of a highly efficient method for generating terahertz waves. Our technique is based on ...

Construction and Characterization of External Cavity Diode Lasers for Atomic Physics

外腔半导体激光器的原子物理的构建与鉴定

Since their development in the late 1980s, cheap, reliable external cavity diode lasers (ECDLs) have replaced complex and expensive traditional dye and ...

Electron Channeling Contrast Imaging for Rapid III-V Heteroepitaxial Characterization

电子通道造影的快速III-V异质特征

Misfit dislocations in heteroepitaxial layers of GaP grown on Si(001) substrates are characterized through use of electron channeling contrast imaging ...

Use of a Multi-compartment Dynamic Single Enzyme Phantom for Studies of Hyperpolarized Magnetic Resonance Agents

一个多腔动态单一酶幻影超极化磁共振剂研究的用途

Imaging of hyperpolarized substrates by magnetic resonance shows great clinical promise for assessment of critical biochemical processes in real time. Due ...

Compact Lens-less Digital Holographic Microscope for MEMS Inspection and Characterization

紧凑的镜头少数字全息显微镜的MEMS检测与表征

A micro-electro-mechanical-system (MEMS) is a widely used component in many industries, including energy, biotechnology, medical, communications, and ...

Data Acquisition Protocol for Determining Embedded Sensitivity Functions

数据采集​​协议确定嵌入式灵敏度功能

The effectiveness of many structural health monitoring techniques depends on the placement of sensors and the location of input forces. Algorithms for ...

Protocol of Electrochemical Test and Characterization of Aprotic Li-O2 Battery

Protocol of Electrochemical Test and Characterization of Aprotic Li-O2 Battery

电化学测试的协议及表征非质子锂的O<sub&gt; 2</sub&gt;电池

We demonstrate a method for electrochemical testing of an aprotic Li-O2 battery. An aprotic Li-O2 battery is made of a Li-metal anode, an aprotic ...

Fabrication of Nanopillar-Based Split Ring Resonators for Displacement Current Mediated Resonances in Terahertz Metamaterials

基于纳米柱裂环谐振器的研制为位移电流共振介导太赫兹超材料

Terahertz (THz) split ring resonator (SRR) metamaterials (MMs) has been studied for gas, chemical, and biomolecular sensing applications because the SRR ...

Guidelines and Experience Using Imaging Biomarker Explorer (IBEX) for Radiomics

Guidelines and Experience Using Imaging Biomarker Explorer IBEX for Radiomics

使用成像生物标志物探险家 (野山羊) Radiomics 的指南和经验

Imaging Biomarker Explorer (IBEX) is an open-source tool for medical imaging radiomics work. The purpose of this paper is to describe how to use IBEX's ...