目的是评估组织固定质量对手术病理免疫组化染色和DNA降解的影响。分析了25个非小细胞肺癌切除标本。切除后，所有肿瘤都按照我们中心的协议进行处理。在血红蛋白和嗜酸性染料（H&E）染色的组织玻片中，根据基底膜剥离，区分了足够和不足够固定的肿瘤区域。在10个免疫组织化学染色中（ALK克隆5A4，PD-L克隆22C3，CAM5.2，CK7，c-Met，KER-MNF116，NapsinA，p40，ROS1，TTF1），确定了足够和不足够固定以及坏死肿瘤区域的免疫反应性H-分数。从相同的区域分离DNA，并测量以碱基对（bp）为单位的DNA断裂。H-分数在H&E足够固定的肿瘤区域中，IHC染色KER-MNF116（H-分数256 vs 15，p = 0.001）和p40（H-分数293 vs 248，p = 0.028）显著高于其他染色。所有其他染色均显示出向足够固定区域更高的免疫反应性的趋势。独立于H&E的足够或不足够固定区域，在肿瘤内所有IHC染色都显示出明显的不同免疫组织化学染色强度，表明其免疫反应性不均匀（H-分数：PD-L1 123 vs 6，p = 0.001；CAM5.2 242 vs 101，p = <0.001；CK7 242 vs 128，p = <0.001；c-MET 99 vs 20，p = <0.001；KER-MNF116 281 vs 120，p = <0.001；Napsin A 268 vs 130，p = 0.005；p40 292 vs 166，p = 0.008；TTF1 199 vs 63，p = <0.001）。无论是否固定充分，DNA片段很少超过300 bp的长度。然而，在固定时间短（<6小时 vs> 16小时）和固定时间短（<24小时 vs> 24小时）的肿瘤中，300 bp和400 bp的DNA片段浓度较高。组织固定不良会导致切除的肺肿瘤某些部位的IHC染色强度降低。这可能会影响IHC分析的可靠性。 版权所有2023。由Elsevier B.V.出版。

The objective is to assess the impact of the quality of tissue fixation in surgical pathology on immunohistochemical (IHC) staining and DNA degradation.Twenty-five non-small cell lung cancer (NSCLC) resection specimens were analyzed. After resection, all tumors were processed according to the protocols in our center. In haematoxylin and eosin (H&E) stained tissue slides, adequately- and inadequately fixed tumor areas were microscopically demarcated, based on basement membrane detachment. In 10 IHC stains ALK (clone 5A4), PD-L (clone 22C3), CAM5.2, CK7, c-Met, KER-MNF116, NapsinA, p40, ROS1, TTF1) the immunoreactivity in H-scores was determined in adequately- and inadequately fixed, and necrotic tumor areas. From the same areas DNA was isolated, and DNA fragmentation in base pairs (bp) was measured.H-scores were significantly higher in H&E adequately fixed tumor areas in IHC stains KER-MNF116 (H-score 256 vs 15, p=0.001) and p40 (H-score 293 vs 248, p=0.028). All other stains showed a trend towards higher immunoreactivity in H&E adequately fixed areas. Independent of H&E adequatelty- or inadequately fixed areas, all IHC stains showed significant different IHC staining intensity within tumors, suggesting heterogeneous immunoreactivity (H-scores: PD-L1 123 vs 6, p = 0.001; CAM5.2 242 vs 101, p=<0.001; CK7 242 vs 128, p=<0.001; c-MET 99 vs 20, p=<0.001; KER-MNF116 281 vs 120, p=<0.001; Napsin A 268 vs 130, p = 0.005; p40 292 vs 166, p = 0.008; TTF1 199 vs 63, p=<0.001). DNA fragments rarely exceeded a length of 300 bp, independent of adequate fixation. However, DNA fragments of 300 and 400 bp had higher concentrations in tumors with short fixation delay (<6 h vs >16 h) and short fixation time (<24 h vs >24 h).Impaired tissue fixation of resected lung tumors results in decreased IHC staining intensity in some parts of the tumor. This may impact the reliability of IHC analysis.Copyright © 2023. Published by Elsevier B.V.