安全注意事项和操作步骤在（A）BSL-4实验室：4.医疗成像过程
Summary
Here, we present an overview of the preparation and animal handling procedures required to safely perform medical imaging in an animal biosafety level 4 laboratory. Computed tomography of a mock-infected guinea pig illustrates these procedures that may be used to evaluate the disease caused by a high consequence pathogen.
Abstract
Medical imaging using animal models for human diseases has been utilized for decades; however, until recently, medical imaging of diseases induced by high-consequence pathogens has not been possible. In 2014, the National Institutes of Health, National Institute of Allergy and Infectious Diseases, Integrated Research Facility at Fort Detrick opened an Animal Biosafety Level 4 (ABSL-4) facility to assess the clinical course and pathology of infectious diseases in experimentally infected animals. Multiple imaging modalities including computed tomography (CT), magnetic resonance imaging, positron emission tomography, and single photon emission computed tomography are available to researchers for these evaluations. The focus of this article is to describe the workflow for safely obtaining a CT image of a live guinea pig in an ABSL-4 facility. These procedures include animal handling, anesthesia, and preparing and monitoring the animal until recovery from sedation. We will also discuss preparing the imaging equipment, performing quality checks, communication methods from “hot side” (containing pathogens) to “cold side,” and moving the animal from the holding room to the imaging suite.
Introduction
国家过敏和传染病研究所（NIAID）在德特里克堡弗雷德里克MD（IRF-弗雷德里克）综合研究机构的使命是执行新发传染性疾病的研究，以了解与严重程度相关的临床疾病过程微生物引起的疾病。该IRF-弗雷德里克必须在ABSL-4实验室1执行的后果严重病原体的动物模型医学成像的独特能力。提供给研究者的成像模态包括：计算机断层扫描（CT），磁共振成像（MRI），正电子发射断层扫描（PET），单光子计算机断层摄影（SPECT），超声波，X射线，和透视。研究人员利用现有的成像能力，监测病情进展和评价干预措施，例如药物治疗和接种疫苗的疗效，在纵向研究。
在IRF-弗雷德里克的成像方式是专门旨在保持设备的核心部件高围堵2,3之外，为维护和修理访问。这种设计成像套件为“热”（含病原体）和分离“冷端”。实现这种分离，专门设计的管构造成高度封闭空间延伸到每一个成像模态（ 图1）的孔中。除了提供生物围堵，这些管保护用于净化高封闭实验室气体和化学品的影像设备。成像的科学家和技术专家从“冷端”操作扫描仪，而比较医学（CM）的工作人员把手，显示器上的“偏热”的动物。由于CM的工作人员必须用影像的科学家紧密合作，协调这些实验中，这种分离可能造成通信的挑战。
评估可供选择之后，CM工作人员出来配有蓝牙耳枚发射短波长的超高频无线电波用来调用成像员工围堵之外的手机。由于设施的设计，无线接入点必须被安装在每个房间克服造成之间的“热”水泥和钢的各层和“冷侧”的信号的干扰。因此，CM的工作人员穿着嘈杂的正压力服和成像人员高度封闭外之间的通信现在是可靠的。摄像机也已安装上的成像室偏热成像人员看到了“偏热”的活动。随着摄像头，成像人员可以指导CM技师动物定位或在最后一分钟改变成像协议。
在IRF-弗雷德里克ABSL-4实验室套装的所有工作人员需要佩戴正压封装服4。穿着这些衣服减少流动性，和重晚点¯x手套附西装加手套多达三个附加层妥协灵巧。其结果是，过程需要更长的时间才能完成，需要精细运动技能任务要困难得多。作为生物安全水平的提高，动物处理和操纵变得更具挑战性和耗时的，特别是与小动物。在ABSL-4实验室程序最多可能需要2-3倍的时间比ABSL-2实验室。
本文的目的是为了在视觉上展示与成像动物模型中使用豚鼠，例如CT扫描过程的ABSL-4环境相关的挑战。
Protocol
Representative Results
Discussion
在本系列前面的文章中都强调了广泛的培训，注重细节，安全程序，并以最高防护实验室12,13安全工作所需的额外工程控制。执行安全工作就是在这些实验室的最高优先级。这一理念更是用活的动物时重要，因为额外的危害，例如潜在的受感染的动物造成咬伤或抓伤或产生气溶胶7。这些程序强调动物的安全装卸和运输从保持房间成像室。麻醉期间动物操作和监测表明，得到观看者…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
The content of this publication does not necessarily reflect the views or policies of the US Department of Health and Human Services (DHHS) or of the institutions and companies affiliated with the authors. This work was funded in part through Battelle Memorial Institute’s prime contract with the US National Institute of Allergy and Infectious Diseases (NIAID) under Contract No. HHSN272200700016I. M.R.H., K.J., D.P., L.B., and J.W. performed this work as employees of Battelle Memorial Institute. Subcontractors to Battelle Memorial Institute who performed this work are: R.B., an employee of Charles River Laboratories – Insourcing Solutions; L.K. and M.R.L., employees of MEDRelief Staffing Inc.; M.G.L. as an employee of Lovelace Respiratory Research Institute, Inc.; and J.H.K. as an employee of Tunnell Government Services, Inc.
Materials
| Micro-Chem Plus | National Chemical Laboratories | 255 | |
| CT scanner | Philips Healthcare | ||
| CT phantom | Philips Healthcare | ||
| Isovue-300 (CT contrast reagent) | Bracco Diagnostics | NDC 0270-1315-30 | |
| Ventilated rack | Lab Products | ||
| Micro-isolator cage | Lab Products | ||
| Biosafety cabinet | Nuaire | ||
| Anesthesia machine | SurgiVet | WWV9000 | |
| Anesthesia induction box | VetEquip | ||
| Anesthesia mask | Henry Schein | ||
| Isoflurane | Henry Schein | ||
| Waste gas scavenging canister | Fisher | F/AIR | |
| Holding cushion | |||
| Ophthalmic ointment | |||
| Vital signs monitor | Bionet | BM3Vet | |
| Mobile phone | Spectralink | 8440 | |
| Blue Tooth ear piece | |||
| Wireless access points | |||
| Sperian positive-pressure suit | Honeywell Safety Products | BSL 4-2 | |
| Outer suit gloves (latex, Ansell Canners and Handlers) | Fisher | 19-019-601 | |
| Outer suit gloves (nitrile/rubber, MAPA) | Fisher | 2MYU1 | |
| Scrubs | Cintas | 60975/60976 | |
| Socks | Cintas | 944 | |
| Duct tape | Pack-N-Tape | 51131069695 | |
| Towels | Cintas | 2720 | |
| Zip lube | Amazon | B000GKBEJA |
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