INVESTIGADORES
KASS Laura
artículos
Título:
Detection of bromodeoxyuridine in formalin-fixed tissue. DNA denaturation following microwave or enzymatic digestion pretreatment is required
Autor/es:
L KASS; J VARAYOUD; HH ORTEGA; M MUÑOZ-DE-TORO; EH LUQUE
Revista:
EUROPEAN JOURNAL OF HISTOCHEMISTRY
Editorial:
SOC ITALIANA ISTOCHIMICA
Referencias:
Año: 2000 vol. 44 p. 185 - 191
ISSN:
1121-760X
Resumen:
The present study was designed to assess the influence of antigen retrieval and/or DNA denaturation on the quantitative estimation of bromodeoxyuridine (BrdU) in formalin-fixed paraffin-embedded tissue. Specimens of small intestine from rats injected with BrdU were routinely fixed and embedded in paraffin. For antigen retrieval, sections were pretreated with microwave oven or enzymatically (pepsin or trypsin). Acid hydrolysis was used as a DNA denaturation method. Immunostaining of BrdU-labeled cells was performed. The best results, regarding tissue morphology and immunostaining, were obtained with microwave pretreatment followed by acid hydrolysis. Enzymatic pretreatment resulted in damage of tissue morphology and/or high background staining. Microwave alone, without DNA denaturation, results in a lower percentage of BrdU positive cells. The significance of validation studies is emphasized when the level of expression of a prognostic marker, such as BrdU, is assessed.
