Immunohistochemistry Quantification by a Digital Computer-Assisted Method Compared to Semiquantitative Analysis

Abstract

PURPOSE: To compare immunostaining quantification obtained by a digital computer-assisted method with the well-established semiquantitative analysis. METHODS: Cytoplasmic staining of galectin-3 was obtained by standard immunohistochemical reactions in 25 cases of welldifferentiated thyroid carcinoma. The expression index that associates the conventional area fraction of labeled cells with the immunostaining intensity score based on visual qualitative observation was used as the semiquantitative analysis. A digital computerassisted method is described based on the use of an image processing program (ImageLab ®). Three parameters were obtained: (1) percentage of labeled cells; (2) digital immunostaining intensity, and (3) digital expression index. The proposed method allows numerical analysis of the immunostaining intensity. RESULTS: There was a strong correlation between the immunostaining intensity obtained by the two methods (Pearson correlation coefficient, r = 0.71, P = 0.0001). The same was observed between expression indexes (Pearson correlation coefficient, r = 0.66, P = 0.0001). CONCLUSION: Results obtained with our proposed digital computer-assisted method for immunoexpression analysis were concordant with the semiquantitative analysis. In addition, digital values can also resolve disagreement among different observers about the quality of staining intensity because the digital method does not classify the results into groups, but rather provides a numerical value for each individual case; thus, it increases the diagnostic and, more importantly, the prognostic sensitivity of the immunohistochemical analysis.

Insertion of the LINE-1 element in the C-MYC gene and immunoreactivity of C-MYC, p53, p21 and p27 proteins in different morphological patterns of the canine TVT

“…Considering the criterion of immunolabeling intensity, in which the fraction of the labeled cell area is evaluated (Matos et al, 2006), it is possible to state that C-MYC, p53, p21, and p27 proteins are present in all TVT samples. In this context, despite the variation of immunolabeling intensity among proteins, it was subtle among the different morphological patterns of TVT.…”

“…It also enables the indirect recognition of the possible mechanisms involved in the growth and progression of neoplasia. Matos et al (2006) and Trompieri-Silveira et al (2009) also adopted the immunolabeling percentage parameter to quantify immunolabeled cells for different proteins. In that sense, Trompieri-Silva et al (2009), considering the percentage of labeled cells, compared the variation of the immunolabeling of the anti-CD3 antibody among the progression and regression phases of TVT, and there were differences in the amount of T lymphocytes in those phases.…”

Distribution and quantification of β-3 adrenergic receptor in tissues of sheep

“…This discrepancy may have been a consequence of the immuno-histochemistry technique. Though it is widely used in basic research for understanding the distribution and localization of biomarkers and differentially expressed proteins in biological tissues, its quantification by a digital computer-assisted method was regarded as semi-quantitative (Matos et al, 2006) and thus not accurate enough. If a post-transcriptional modification does affect the protein expression deserves further study.…”

Immunohistochemical Expression of Cyclin D1 in Invasive Ductal Carcinoma of Human Breast

“…Maximum, minimum and integrity of intense color based on Gray-level acquisition and analysis of the data level were carried out by reading spot fixed areas of 10 images for each case. The IOD based on Gray-level transition probabilities in digitized images was graded from light to dark (0 up to 250) [16]. The average score across the whole image should be taken; IOD in digitized images was calibrated from strong to light (180 down to 70) by pixel.…”