Quantification of immunohistochemistry—issues concerning methods, utility and semiquantitative assessment II

Abstract

Immunohistochemistry is entering its fourth decade of use on formalin-fixed paraffin-embedded tissues. Over this period the method has evolved to become a major part of the practice of diagnostic surgical pathology worldwide. From the beginning immunohistochemistry has been adapted to provide a range of markers of cell lineage and tissue type, with particular application to the diagnosis and classification of tumours. In this modality immunohistochemical methods were employed simply as 'special stains', the results of which were evaluated qualitatively by the pathologist, as for any other stain. More recently, attention has shifted to the demonstration of prognostic markers in tumour cells, driven by the advent of molecular biology and the discovery of numerous regulatory molecules, coupled with manufacture of the corresponding specific antibodies. Immunohistochemistry has rapidly adapted to this new use, but in so doing the demand for quantification has become paramount; it is no longer enough that the 'stain' is there; rather it is a question of 'How much is there?'. This review explores the limitations of immunohistochemistry when employed in a semiquantitative mode, and explores the possibility of fulfilling the full potential of immunohistochemistry, as a true quantitative immunoassay applied in a tissue section environment.

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

“…There is debate among pathologists as to where the absolute ''sweet spot'' is for optimal section thickness. Taylor and Levenson (2006) reported that this ''sweet spot'' was approximately 5 mm and that ''that uniform preparation of FFPE sections that are less than 5 mm in thickness is not possible.'' In contrast, the recommended thickness for performing the Hercept test is 4 mm; and for many pharmas (including ours), the standard thickness of a histologic section is 3 mm (Wolff et al 2007) (Figure 4).…”

Prenatal cocaine exposure enhances responsivity of locus coeruleus norepinephrine neurons: Role of autoreceptors

“…Briefly, one of the five sets of tissue was used, consisting of 40 µm sections taken every 200 µm starting rostral to the LC and continuing past the caudal edge of the LC. Immunohistochemical staining was carried out in 2 batches using identical methods, timing, reagents, buffers, and antibodies over a 2-week period to minimize any potential differences (Taylor and Levenson, 2006). Each batch contained tissues from half of the rats in each treatment group, and they were coded to blind the investigators.…”

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].…”