Casadonte Rita, Kriegsmann Jörg, Kriegsmann Mark, Kriegsmann Katharina, Torcasio Roberta, Gallo Cantafio Maria Eugenia, Viglietto Giuseppe, Amodio Nicola
Proteopath GmbH, 54296 Trier, Germany.
Department of Medicine, Faculty of Medicine and Dentistry, Danube Private University, 3500 Krems, Austria.
Cancers (Basel). 2023 Feb 3;15(3):974. doi: 10.3390/cancers15030974.
Sample processing of formalin-fixed specimens constitutes a major challenge in molecular profiling efforts. Pre-analytical factors such as fixative temperature, dehydration, and embedding media affect downstream analysis, generating data dependent on technical processing rather than disease state. In this study, we investigated two different sample processing methods, including the use of the cytospin sample preparation and automated sample processing apparatuses for proteomic analysis of multiple myeloma (MM) cell lines using imaging mass spectrometry (IMS). In addition, two sample-embedding instruments using different reagents and processing times were considered. Three MM cell lines fixed in 4% paraformaldehyde were either directly centrifuged onto glass slides using cytospin preparation techniques or processed to create paraffin-embedded specimens with an automatic tissue processor, and further cut onto glass slides for IMS analysis. The number of peaks obtained from paraffin-embedded samples was comparable between the two different sample processing instruments. Interestingly, spectra profiles showed enhanced ion yield in cytospin compared to paraffin-embedded samples along with high reproducibility compared to the sample replicate.
福尔马林固定标本的样本处理是分子分析工作中的一项重大挑战。诸如固定剂温度、脱水和包埋介质等分析前因素会影响下游分析，产生依赖于技术处理而非疾病状态的数据。在本研究中，我们研究了两种不同的样本处理方法，包括使用细胞离心涂片机制备样本以及使用自动样本处理设备，通过成像质谱（IMS）对多发性骨髓瘤（MM）细胞系进行蛋白质组分析。此外，还考虑了两种使用不同试剂和处理时间的样本包埋仪器。将固定在4%多聚甲醛中的三种MM细胞系，要么使用细胞离心涂片制备技术直接离心到载玻片上，要么用自动组织处理仪处理制成石蜡包埋标本，再进一步切成载玻片用于IMS分析。从石蜡包埋样本中获得的峰数在两种不同的样本处理仪器之间具有可比性。有趣的是，光谱图谱显示，与石蜡包埋样本相比，细胞离心涂片的离子产率更高，并且与样本重复相比具有更高的重现性。
