Interpretation and Quantification of Immunostains

Abstract

Despite the fact that immunohistochemistry is widely used in routine diagnostic work and is a very common part of scientific reports in pathology and cytology, its standardization still lags behind. Interpretation of immunostains should be based on microanatomic distribution of the staining, proportion of positively stained cells, staining intensity, if relevant, and cutoff levels. These parameters should be shown to be reasonably reproducible and should be clearly defined in publications. Uniformity in the setting of thresholds could probably benefit from interlaboratory control materials containing defined amounts of the target antigen. Reliable and precise quantitative immunohistochemistry requires the use of control materials containing defined amounts of the target antigen and processed alongside the specimen combined with automated computer-assisted microspectrophotometry. Application of these suggestions is hoped to improve standardization and to facilitate communication in the field of immunohistochemistry.

Improving evaluation of the distribution and density of immunostained cells in breast cancer using computerized video image analysis

“…This may potentially lead to 'dilution' of the available data. High intra-observer variability and difficulties with reproducibility are well recognized as serious limitations of standard manual quantal visual cell density grading methods, [2][3][4] and other methods have been diligently sought in an attempt to reduce or eliminate these types of errors and problems.…”

“…2 However, a lack of repeatability (due to significant inter-and intra-observer variability) has proved to be a major limitation with such methodologies. [1][2][3][4][5] Moreover, visual quantitation is relatively time-intensive, has some degree of imprecision and requires a certain level of experience. 6,7 The difficulties inherent with standard quantal visual scale methods relate to the accurate placement of a particular density of immunostaining into a specific category, as this process is relatively subjective and requires a number of assumptions.…”

“…[2][3][4][5] In reality, cell density is a continuous biological spectrum that ranges from zero, in which there are no immunostained cells, to maximal, in which there are densely packed contiguous immunostained cells. Therefore, the advent of novel computer-assisted video image analysis methods (VIA) is potentially significant, as it provides the ability to quantitate cells using a continuous scale from zero to maximal density, rather than a quantal or discontinuous grading scale as determined by standard visual methods.…”

Glioblastoma pediátrico: estudo clínico patológico de 12 casos com imunoistoquímica para proteína p53

“…Em cada um dos casos foram contadas 2610 células (5 a 10 CGA). As áreas selecionadas foram as mais representativas, com maior concentração do corante, chamadas de "hot spots" [14][15][16] .…”

Quantitative Assessment and Classification of Tissue-Based Biomarker Expression With Color Content Analysis

“…34 It has been proposed that computer-aided quantification of expression would be a useful tool for reducing interlaboratory and interobserver variability as well as for increasing throughput and reducing pathologist workload. [35][36][37][38] In the case of HER2/neu, the College of American Pathologists/American Society of Clinical Oncology (CAP/ASCO) guidelines recommend image analysis as an effective tool for achieving consistent interpretation of IHC HER2/neu staining, provided that a pathologist confirms the result. 39 The use of computer aids is also compatible with the increasing use of digital imaging in pathology; the field of digital pathology, both for clinical practice and the development of new biomarkers, has been growing rapidly, driven by the need for high throughput and recent developments in microscopy imaging, such as automated whole-slide imaging systems.…”