The impact of pre-analytical processing on staining quality for H&E, dual hapten, dual color in situ hybridization and fluorescent in situ hybridization assays

The Use of Immunohistochemistry for Biomarker Assessment—Can It Compete with Other Technologies?

“…Even H&E staining has been reported to be compromised with fixation in 10% NBF for 8 hours or less (Babic et al 2010;Dapson 2007).…”

The prognostic and predictive value of excision repair cross-complementation group 1 (ERCC1) protein in 1288 patients with head and neck squamous cell carcinoma treated with platinum-based therapy: a meta-analysis

“…It has been shown that ERCC1 immunodetection faces several methodological challenges including tissue processing, rates of interobserver agreement, geographic variation in protein expression and, most importantly, lack of the standardized cutoff value in semi-quantitative IHC [39][40][41]. It has been also reported that the most frequently used antibody 8F1 has not been specific for ERCC1 [42].…”

The challenge of NSCLC diagnosis and predictive analysis on small samples. Practical approach of a working group

“…Fixation times below 6 h are not recommended, since conventional staining as well as immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH) analysis are adversely affected [55]. Antigen preservation for immunohistochemistry is epitope dependent, and may for some not be hampered by fixation times of up to 120 h [56], but signals in FISH analysis become weaker after fixation exceeding 48 h [55]. Amplifiable RNA can be retrieved from FFPE even after prolonged fixation times of 21 days, though with less efficiency [57].…”