Influence of slide aging on results of translational research studies using immunohistochemistry

Abstract

Several reports have shown that a long delay between cutting sections and immunohistochemical (IHC) staining can decrease the IHC reaction intensity. However, systematic large-scale studies to investigate to what extent this problem may influence the outcome of translational research studies are lacking. In this study, we used a tissue microarray (TMA) approach to investigate the influence of slide age on comparisons between the results of IHC analyses for estrogen receptor (ER), progesterone receptor (PR), cyclin D1, HER2 (HercepTest), and E-cadherin and clinical outcome in a series of 522 breast cancer patients. Old TMA sections stored for 6 months at 41C and freshly cut sections were analyzed under exactly identical experimental conditions. As compared to results obtained on freshly cut sections, the frequency of positivity on old sections decreased from 65 to 46% for ER (Po0.0001), from 33 to 18.5% for PR (Po0.0001), from 16.3 to 9.6% for HER2 (P ¼ 0.0047), from 45.1 to 37.7% for cyclin D1 (P ¼ 0.10), and from 58.9 to 32.9% for E-cadherin (Po0.0001). Despite the lower fraction of positive cases, most associations between IHC data and tumor phenotype that were observed in fresh section analysis were also found when old section data were analyzed. The results confirm that slide aging has a great influence on the intensity of IHC staining in individual cases, but they also suggest that many clinicopathological associations can be detected if suboptimally processed sections are used for IHC.

Effects of Long-Term Storage on the Detection of Proteins, DNA, and mRNA in Tissue Microarray Slides

“…That study confirms that the Envision system used in our study is one of the most sensitive systems presently used and is significantly more sensitive than the older detection systems used in earlier studies. The importance of the detection system, as well as the AR procedure, for preserved antigenicity is thus obvious in our study compared with older studies (Fergenbaum et al 2004;Mirlacher et al 2004).…”

High throughput tissue microarray analysis of FHIT expression in diffuse large cell B-cell lymphoma from Saudi Arabia

“…PCR based hypermetylation analysis clearly is a more robust and reproducible method than immunohistochemistry, which is prone to numerous technical shortcomings. 48,49 However, our data also raise the possibility that certain FHIT inactivation mechanisms could be linked to different diffuse large B-cell lymphoma subtypes. With the exception of one case, FHIT hypermethylation was only seen in the nongerminal center diffuse large B-cell lymphoma subtype.…”

Frequent homogeneous HER-2 amplification in primary and metastatic adenocarcinoma of the esophagus

“…It is well known, that prolonged formalin fixation can disturb immunohistochemical detectability of HER-2 protein while insufficient formalin fixation with consecutive tissue exposure to ethanol during technical processing can lead to false positive HER-2 IHC. [52][53][54] Considering the encouraging results of clinical trials in breast cancer, it could be speculated that Herceptin s might also represent a possible option for HER-2 amplified esophageal adenocarcinomas. The only published clinical Herceptin s trial for esophagus cancer patients was a phase I study testing 30 patients, including six patients with HER-2 amplified tumors.…”