Department of Biochemistry, Vanderbilt University, Nashville, Tennessee, USA; Mass Spectrometry Research Center, Vanderbilt University, Nashville, Tennessee, USA.
Division of Nephrology and Hypertension, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA.
Kidney Int. 2022 Jan;101(1):137-143. doi: 10.1016/j.kint.2021.08.033. Epub 2021 Oct 5.
The human kidney is composed of many cell types that vary in their abundance and distribution from normal to diseased organ. As these cell types perform unique and essential functions, it is important to confidently label each within a single tissue to accurately assess tissue architecture and microenvironments. Towards this goal, we demonstrate the use of co-detection by indexing (CODEX) multiplexed immunofluorescence for visualizing 23 antigens within the human kidney. Using CODEX, many of the major cell types and substructures, such as collecting ducts, glomeruli, and thick ascending limb, were visualized within a single tissue section. Of these antibodies, 19 were conjugated in-house, demonstrating the flexibility and utility of this approach for studying the human kidney using custom and commercially available antibodies. We performed a pilot study that compared both fresh frozen and formalin-fixed paraffin-embedded healthy non-neoplastic and diabetic nephropathy kidney tissues. The largest cellular differences between the two groups was observed in cells labeled with aquaporin 1, cytokeratin 7, and α-smooth muscle actin. Thus, our data show the power of CODEX multiplexed immunofluorescence for surveying the cellular diversity of the human kidney and the potential for applications within pathology, histology, and building anatomical atlases.
人类肾脏由许多细胞类型组成，这些细胞类型在正常和患病器官中的丰度和分布各不相同。由于这些细胞类型具有独特且必不可少的功能，因此在单个组织中准确地标记每个细胞类型以准确评估组织结构和微环境非常重要。为此，我们展示了使用共检测索引（CODEX）多重免疫荧光技术来可视化人类肾脏中的 23 种抗原。使用 CODEX，可以在单个组织切片中可视化许多主要的细胞类型和亚结构，如集合管、肾小球和厚升支。这些抗体中有 19 种是在内部共轭的，这证明了这种方法的灵活性和实用性，可用于使用定制和市售抗体研究人类肾脏。我们进行了一项初步研究，比较了新鲜冷冻和福尔马林固定石蜡包埋的健康非肿瘤和糖尿病肾病肾脏组织。两组之间最大的细胞差异是在标记有水通道蛋白 1、细胞角蛋白 7 和α-平滑肌肌动蛋白的细胞中观察到的。因此，我们的数据表明 CODEX 多重免疫荧光技术具有调查人类肾脏细胞多样性的强大功能，并且在病理学、组织学和构建解剖图谱方面具有应用潜力。
