title: Immunocytochemistry associated with oral exfoliative cytology: methodological analysis..
abstract: OBJECTIVE
  To evaluate different immunocytochemical protocol variations to find the most effective protocol for the analysis of involucrin, epidermal growth factor receptor (EGFR), and E-cadherin antibodies. Exfoliative cytology is a noninvasive method used to monitor and screen for early changes in the oral mucosa of patients exposed to carcinogens such as tobacco and alcohol. It has been postulated that its association with immunocytochemistry may improve the effectiveness of the screening process.
  
  
  METHODS
  Four graduate students from Porto Alegre in southern Brazil had oral smears collected from the border of the tongue using a cytobrush. The following variables were analyzed: cell membrane permeability, antigen retrieval method (microwave oven or water bath), antibody incubation time (overnight or 1 hour), detection system used (Envision or LSAB), and chromogen incubation time (10 seconds or 5 minutes).
  
  
  RESULTS
  Best results were obtained with the following combinations: (1) for involucrin: water bath, 1-hour incubation for primary antibody, Envision, and chromogen incubation for 10 seconds; (2)for EGFR: microwave, overnight incubation, LSAB, and chromogen incubation for 5 minutes; and (3) for E-cadherin: water bath, over-night incubation, Envision, and chromogen incubation for 5 minutes.
  
  
  CONCLUSIONS
  Our findings suggest that each antibody requires a specific immunocytochemical protocol to guarantee optimal results with oral smears.
authors.forename: Alessandra Dutra
authors.initials: AD
authors.lastname: da Silva
authors.forename: Celina Faig
authors.initials: CF
authors.lastname: Lima
authors.forename: Bruna Jalfim
authors.initials: BJ
authors.lastname: Maraschin
authors.forename: Natália Koerich
authors.initials: NK
authors.lastname: Laureano
authors.forename: Natália Batista
authors.initials: NB
authors.lastname: Daroit
authors.forename: Fernanda
authors.initials: F
authors.lastname: Brochier
authors.forename: Manoel
authors.initials: M
authors.lastname: Sant'Ana Filho
authors.forename: Fernanda
authors.initials: F
authors.lastname: Visioli
authors.forename: Pantelis Varvaki
authors.initials: PV
authors.lastname: Rados
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000032:analysis; D003581:Cytodiagnosis / Q000379:methods* / Q000592:standards*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods* / Q000592:standards*; D008297:Male; D009061:Mouth Mucosa; D009062:Mouth Neoplasms / Q000175:diagnosis; D055815:Young Adult
chemical_list: D014408:Biomarkers, Tumor