Comparative assessment between objective and subjective methods in slides stained by immunohistochemistry.
Objective methods of assessment are often required in scientific studies. Histological tests with immunohistochemical staining can be assessed by photometry.


OBJECTIVE
To compare this objective method with the subjective evaluation performed by three independent examiners, using slides of acquired middle ear cholesteatomas.


METHODS
We selected a total of 54 cholesteatoma images, immunohistochemically stained by anti-TNF-R2 (32 slides) and anti-TGF-α, (22 slides). The secondary antibody used in the two groups was the Max Polymer Detection System (Novo Link Kit, Novocastra®, UK). The samples were processed by a digital slide scanner (ScanScope - Aperio). The selected sites were analyzed by photometry.


RESULTS
The objective assessment by photometry was compared with the subjective evaluation by three examiners and subjected to statistical analysis. The Statistical analysis revealed moderate reproducibility (K values between 0.41 and 0.60) for both groups.


CONCLUSIONS
Our study showed that the irregular characteristics of middle ear cholesteatoma slides stained by immunohistochemistry prevents its proper objective evaluation, while the subjective assessment by experienced examiners was more reliable.
Santa Casa de São Paulo, Faculdade de Ciências Médicas, Departamento de Otorrinolaringologia.
Fernando de Andrade Quintanilha
Fde A
Ribeiro
Santa Casa de São Paulo, Faculdade de Ciências Médicas, Departamento de Morfologia.
Celina Siqueira Barbosa
CS
Pereira
Santa Casa de São Paulo, Faculdade de Ciências Médicas, curso de Medicina.
Ricardo Junchen
RJ
Chi
Santa Casa de São Paulo, Faculdade de Ciências Médicas, curso de Medicina.
Patrícia Lumi
PL
Yokomizo
Hospital do câncer de Barretos, Instituto de ensino e pesquisa, núcleo de apoio ao pesquisador.
José Humberto Tavares Guerreiro
JH
Fregnani
Hospital AC Camargo, Departamento de Anatomia Patológica, São Paulo.
Rafael Malagoli
RM
Rocha
D001323:Autoantibodies / Q000032:analysis*; D018424:Cholesteatoma, Middle Ear / Q000473:pathology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D015588:Observer Variation; D010783:Photometry; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000032:analysis*; D015203:Reproducibility of Results; D016211:Transforming Growth Factor alpha / Q000032:analysis*
D001323:Autoantibodies; D047889:Receptors, Tumor Necrosis Factor, Type II; D016211:Transforming Growth Factor alpha