Fernandes, Diego TetznerVan Heerden, Willem Francois PetrusPrado Ribeiro, Ana CarolinaBrandao, Thais BiancaDe Mello, Evandro SobrozaRivera, CesarVan Heerden, Marlene B.Gondak, RogerioSantos- Silva, Alan RogerVargas, Pablo AgustinLopes, Marcio Ajudarte2020-11-182020-11-182020-05Fernandes, D.T., Van Heerden, W.F.P., Ribeiro, A.C.P. et al. 2020, 'Different methods of cell quantification can lead to different results : a comparison of digital methods using a pilot study of dendritic cells in HIV-positive patients', Medicina Oral, Patologia Oral Y Cirugia Bucal, vol. 25, no. 3, pp. e431-e438.1698-4447 (print)1698-6946 (online)10.4317/medoral.23472http://hdl.handle.net/2263/77074BACKGROUND : Although new digital pathology tools have improved the positive cell quantification, there is a heterogeneity of the quantification methods in the literature. The aim of this study was to evaluate and propose a novel dendritic cells quantification method in squamous cell carcinoma comparing it with a conventional quantification method. MATERIAL AND METHODS : Twenty-six squamous cell carcinomas HIV-positive cases affecting the oropharynx, lips and oral cavity were selected. Immunohistochemistry for CD1a, CD83, and CD207 was performed. The immunohistochemical stains were evaluated by automated examination using a positive pixel count algorithm. A conventional quantification method (unspecific area method; UA) and a novel method (specific area method; SA) were performed obtaining the corresponding density of positive dendritic cells for the intratumoral and peritumoral regions. The Mann-Whitney U test was used to verify the influence of the quantification methods on the positive cell counting according to the evaluated regions. Data were subjected to the ANOVA and Student's t-test to verify the influence of the tumour location, stage, histological grade, and amount of inflammation on the dendritic cells density counting. RESULTS : The cell quantification method affected the dendritic cells counting independently of the evaluated region (P-value <0.05). Significant differences between methods were also observed according to the tumour features evaluations. CONCLUSIONS : The positive cell quantification method influences the dendritic cells density results. Unlike the conventional method (UA method), the novel SA method avoids non-target areas included in the hotspots improving the reliability and reproducibility of the density cell quantification.en© Medicina Oral S. L. C.I.F.Cell countsImmunohistochemistryDendritic cellsHead and neck neoplasmsHuman immunodeficiency virus (HIV)Article