Department of Biological Chemistry, University of California, Irvine, CA 92697, USA.
Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, CA 92697, USA.
Int J Mol Sci. 2023 Apr 10;24(8):7008. doi: 10.3390/ijms24087008.
To image 4-plex immunofluorescence-stained tissue samples at a low cost with cellular level resolution and sensitivity and dynamic range required to detect lowly and highly abundant targets, here we describe a robust, inexpensive (<$9000), 3D printable portable imaging device (Tissue Imager). The Tissue Imager can immediately be deployed on benchtops for in situ protein detection in tissue samples. Applications for this device are broad, ranging from answering basic biological questions to clinical pathology, where immunofluorescence can detect a larger number of markers than the standard H&E or chromogenic immunohistochemistry (CIH) staining, while the low cost also allows usage in classrooms. After characterizing our platform's specificity and sensitivity, we demonstrate imaging of a 4-plex immunology panel in human cutaneous T-cell lymphoma (CTCL) formalin-fixed paraffin-embedded (FFPE) tissue samples. From those images, positive cells were detected using CellProfiler, a popular open-source software package, for tumor marker profiling. We achieved a performance on par with commercial epifluorescence microscopes that are >10 times more expensive than our Tissue Imager. This device enables rapid immunofluorescence detection in tissue sections at a low cost for scientists and clinicians and can provide students with a hands-on experience to understand engineering and instrumentation. We note that for using the Tissue Imager as a medical device in clinical settings, a comprehensive review and approval processes would be required.
为了以低成本、细胞水平分辨率和所需的灵敏度和动态范围对 4 重免疫荧光染色的组织样本进行成像，以检测低丰度和高丰度靶标，我们在这里描述了一种强大、廉价（<$9000）、可 3D 打印的便携式成像设备（组织成像仪）。组织成像仪可以立即部署在台式机上，用于原位检测组织样本中的蛋白质。该设备的应用非常广泛，从回答基本的生物学问题到临床病理学，免疫荧光可以检测比标准 H&E 或显色免疫组化（CIH）染色更多的标记物，而低成本也允许在课堂上使用。在对我们平台的特异性和灵敏度进行了表征之后，我们展示了对人类皮肤 T 细胞淋巴瘤（CTCL）福尔马林固定石蜡包埋（FFPE）组织样本的 4 重免疫学检测。从这些图像中，使用 CellProfiler 检测了阳性细胞，这是一种流行的开源软件包，用于肿瘤标志物分析。我们的性能与商业荧光显微镜相当，而商业荧光显微镜的价格是我们 Tissue Imager 的 10 倍以上。这种设备能够以低成本在组织切片中进行快速免疫荧光检测，为科学家和临床医生提供了一种实践经验，让学生了解工程和仪器仪表。我们注意到，要将 Tissue Imager 用作临床环境中的医疗设备，需要进行全面的审查和批准程序。
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