title: Methods of cytologic smear preparation and fixation. Effect on the immunoreactivity of commonly used anticytokeratin antibody AE1/AE3.
abstract: OBJECTIVE
  To evaluate the effect of fixation and methods of cytologic smear preparation on the immunoreactivity of commonly used anticytokeratin antibody AE1/AE3.
  
  
  METHODS
  Scrape cytology smears and formalin-fixed, paraffin-embedded tissue sections (FPTS) of 20 unfixed, fresh specimens submitted for intraoperative consultation were studied by the immunoperoxidase method. In addition to the morphologic examination, the smears and FPTS were evaluated for intensity and proportion scores. For each specimen, two scrape cytology smears were wet fixed in 95% ethanol, and 12 smears were air dried without fixation. Air-dried smears were either postfixed after rehydration in saline or fixed directly without rehydration by one of the three fixatives: alcoholic formalin, 95% ethanol with 5% acetic acid or 95% ethanol.
  
  
  RESULTS
  Both intensity and proportion scores were higher with rehydrated, air-dried smears as compared to those without rehydration and were comparable to those with wet-fixed smears and FPTS. In the rehydrated group, the optimum results were achieved when the smears were postfixed with alcoholic formalin.
  
  
  CONCLUSIONS
  The method of preparation and fixation had variable effects on the immunoreactivity of anticytokeratin antibody AE1/AE3. The optimum results were achieved with saline-rehydrated, air-dried smears post-fixed in alcoholic formalin. To evaluate the role of inter-sample variation, further, larger studies are recommended on this and other antibodies before applying them to different types of cytologic smears.
authors: Department of Pathology, Medical College of Wisconsin, 9200 West Wisconsin Avenue, Milwaukee, Wisconsin 53226, USA. vshidham@mcw.edu
authors: V B
authors: VB
authors: Shidham
authors: P F
authors: PF
authors: Lindholm
authors: A
authors: A
authors: Kajdacsy-Balla
authors: C C
authors: CC
authors: Chang
authors: R
authors: R
authors: Komorowski
mesh_terms: D000906:Antibodies; D001707:Biopsy, Needle; D006801:Humans; D007124:Immunoenzyme Techniques; D007633:Keratins / Q000276:immunology / Q000378:metabolism*; D009369:Neoplasms / Q000473:pathology; D016707:Tissue Fixation
chemical_list: D000906:Antibodies; D007633:Keratins