Author/Editor     Pinheiro, C.; Roque, R.; Adriano, A.; Mendes, P.; Praça, M.; Reis, I.; Pereira, T.; Srebotnik-Kirbiš, Irena; André, S.
Title     Optimization of immunocytochemistry in cytology
Type     članek
Vol. and No.     Letnik 26, št. 1
Publication year     2015
Volume     str. 38-43
ISSN     0956-5507 - Cytopathology : official journal of the British Society for Clinical Cytology
Language     eng
Abstract     Objective: A new protocol for fixation and slide preservation was evaluated in order to improve the quality of immunocytochemical reactions on cytology slides. Methods: The quality of immunoreactions was evaluated retrospectively on 186 cytology slides (130 direct smears, 56 cytospins) prepared from different cytology samples. Ninety-three of the slides were air dried, stored at -20 ⁰C and fixed in acetone for 10 minutes (Protocol 1), whereas the other 93 were immediately fixed in methanol at -20 ⁰C for at least 30 minutes, subsequently protected with polyethylene glycol (PEG) and stored at room temperature (Protocol 2). Immunocytochemical staining, with eight primary antibodies, was performed on a Ventana BenchMark Ultra instrument using an UltraView Universal DAB Detection Kit. The following parameters were evaluated for each immunoreaction: morphology preservation, intensity of specific staining, background and counterstain. The slides were blinded and independently scored by four observers with marks from 0 to 20. Results: The quality of immunoreactions was better on methanol-fixed slides protected with PEG than on air-dried slides stored in the freezer: X = 14.44 % 3.58 versus X = 11.02 % 3.86, respectively (P < 0.001). Conclusion: Immediate fixation of cytology slides in cold methanol with subsequent application of PEG is an easy and straightforward procedure that improves the quality of immunocytochemical reactions and allows the storage of the slides at room temperature.
Keywords     cytology
immunocytochemistry
polyethylene glycol
citologija
imunocitokemija
polietilen glikol