Translational Ophthalmic Pathology Platform, Singapore Eye Research Institute, Singapore, Singapore.
A. Menarini Biomarkers Singapore Pte Ltd, Singapore.
Ocul Immunol Inflamm. 2021 Apr 3;29(3):430-439. doi: 10.1080/09273948.2019.1636093. Epub 2019 Aug 16.
Vitreoretinal lymphoma (VRL) is a potentially fatal intraocular malignancy. Diagnosis is hampered by poor preservation of morphology and DNA/RNA integrity, which precludes adjunctive molecular analysis. We aimed to determine the optimum fixative protocol for VRL biopsies that permits cytology, IHC/flow cytometry and molecular analyses. Six fixatives were compared on cultured Pfeiffer cells used as a cellular model. Cells were fixed and evaluated on cellular morphology, antibody staining, DNA/RNA amount and integrity. VRL clinical cases were used as validation and proof-of-concept. PreservCyt was the best fixative for preserving cellular morphology and high-quality RNA/DNA from vitreous fluid biopsies. Cells from clinical VRL cases fixed with PreservCyt showed adequate cellular morphology and IHC positivity. Sufficient DNA was obtained for IgH clonality and MYD88 mutation detection using remnant cytological fluid. PreservCyt maintains good morphology and RNA/DNA integrity suggesting that it is a suitable fixative for VRL diagnosis and molecular analysis.
玻璃体内视网膜淋巴瘤 (Vitreoretinal lymphoma, VRL) 是一种潜在致命的眼内恶性肿瘤。由于形态学和 DNA/RNA 完整性保存不佳，诊断受到阻碍，无法进行辅助分子分析。我们旨在确定最适合 VRL 活检的固定剂方案，以允许细胞学、免疫组化/流式细胞术和分子分析。我们比较了六种固定剂对作为细胞模型的培养 Pfeiffer 细胞的影响。评估细胞固定后的细胞形态、抗体染色、DNA/RNA 含量和完整性。使用 VRL 临床病例进行验证和概念验证。在保留细胞形态和高质量 RNA/DNA 方面，PreservCyt 是最好的固定剂。来自临床 VRL 病例的细胞用 PreservCyt 固定后，显示出足够的细胞形态和免疫组化阳性。从剩余细胞学液体中获得了足够的 DNA ，用于 IgH 克隆性和 MYD88 突变检测。PreservCyt 保持良好的形态和 RNA/DNA 完整性，表明它是 VRL 诊断和分子分析的合适固定剂。
