Faculty of Medicine, Eye Center, University of Freiburg, Killianstrasse 5, 79106, Freiburg, Germany.
Institute of Anatomy, Wuerzburg University, Wuerzburg, Germany.
BMC Ophthalmol. 2021 Sep 20;21(1):338. doi: 10.1186/s12886-021-02099-8.
Imaging mass cytometry (IMC) combines the principles of flow cytometry and mass spectrometry (MS) with laser scanning spatial resolution and offers unique advantages for the analysis of tissue samples in unprecedented detail. In contrast to conventional immunohistochemistry, which is limited in its application by the number of possible fluorochrome combinations, IMC uses isoptope-coupled antibodies that allow multiplex analysis of up to 40 markers in the same tissue section simultaneously.
In this report we use IMC to analyze formalin-fixed, paraffin-embedded conjunctival tissue. We performed a 18-biomarkers IMC analysis of conjunctival tissue to determine and summarize the possibilities, relevance and limitations of IMC for deciphering the biology and pathology of ocular diseases.
Without modifying the manufacturer's protocol, we observed positive and plausible staining for 12 of 18 biomarkers. Subsequent bioinformatical single-cell analysis and phenograph clustering identified 24 different cellular clusters with distinct expression profiles with respect to the markers used.
IMC enables highly multiplexed imaging of ocular samples at subcellular resolution. IMC is an innovative and feasible method, providing new insights into ocular disease pathogenesis that will be valuable for basic research, drug discovery and clinical diagnostics.
成像质谱流式细胞术（IMC）结合了流式细胞术和质谱（MS）的原理，具有激光扫描空间分辨率，为以前所未有的细节分析组织样本提供了独特的优势。与传统的免疫组织化学相比，免疫组织化学受到可能的荧光染料组合数量的限制，而 IMC 则使用等光耦连抗体，允许在同一切片上同时对多达 40 个标记物进行多重分析。
在本报告中，我们使用 IMC 分析了福尔马林固定、石蜡包埋的结膜组织。我们对结膜组织进行了 18 种生物标志物的 IMC 分析，以确定和总结 IMC 用于破译眼部疾病生物学和病理学的可能性、相关性和局限性。
在不修改制造商方案的情况下，我们观察到 18 种生物标志物中的 12 种具有阳性和合理的染色。随后的单细胞生物信息学分析和表型聚类确定了 24 个不同的细胞簇，这些细胞簇在使用的标记物方面具有不同的表达谱。
IMC 能够以亚细胞分辨率对眼部样本进行高度多重成像。IMC 是一种创新且可行的方法，为眼部疾病发病机制提供了新的见解，这将对基础研究、药物发现和临床诊断具有重要价值。
