Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.21/3846
Título: Optimization of immunocytochemistry in cytology: comparison of two protocols for fixation and preservation on cytospin and smear preparations
Autor: Pinheiro, C.
Roque, Rúben
Adriano, A.
Mendes, P.
Praça, M.
Reis, I.
Pereira, T.
Srebotnik Kirbis, I.
André, S.
Palavras-chave: Cytology
Fixation
Immunocytochemistry
Polyethylene glycol
Slide storage
Data: Mai-2014
Editora: Wiley
Citação: Pinheiro C, Roque R, Adriano A, Mendes P, Praça M, Reis I, et al. Optimization of immunocytochemistry in cytology: comparison of two protocols for fixation and preservation on cytospin and smear preparations. Cytopathology. 2014 May 19. [Epub ahead of print]
Resumo: 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 °C and fixed in acetone for 10 minutes (Protocol 1), whereas the other 93 were immediately fixed in methanol at -20 °C 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.
Peer review: yes
URI: http://hdl.handle.net/10400.21/3846
DOI: 10.1111/cyt.12156
Versão do Editor: http://onlinelibrary.wiley.com/doi/10.1111/cyt.12156/abstract
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