Applying immunohistochemistry to alcohol-fixed paraffin-embedded tissues: an innovative technique to reduce use of formaldehyde

作者：Panzacchi Simona; Boiani Sara; Mandrioli Daniele; Piccioli Milena; Belpoggi Fiorella^*

来源：European Journal of Oncology, 2013, 18(2): 75-83.

• alcohol-fixation
• IHC
• AFPE post-fixation
• FFPE
• formalin fixation

Recent advances in molecular biology and pathology have opened new opportunities for refining our knowledge of pathophysiologic events and biomarkers. Particular interest in applying these novel methods to current and archived tissues collected in experimental and epidemiological/clinical studies is evident. Until now, it has not always been possible to use archived alcohol-fixed paraffin-embedded (AFPE) tissues for immunohistochemisty (IHC), because AFPE slices and blocks were not often amenable to standard IHC methods. In order to solve this problem, we developed a simple method of post-fixation, which allows to use, on AFPE slices, standard IHC protocols already used for formalin-fixed paraffin-embedded (FFPE) samples. For the assessment of post-fixation processing and to test the feasibility of IHC, we selected the spleen from Sprague-Dawley rats as a demonstrative tissue. Antibodies to PAX5, CD3, CD68 and Ki-67, were tested on FFPE, AFPE and AFPE post-fixed spleen samples. The specificity of antibodies to bind different epitopes expressed in spleen tissue was maintained in FFPE and AFPE post-fixed sections, according to anatomical localization. Post-fixation of AFPE samples did not affect tissue morphology and IHC results were comparable to the FFPE sections in terms of sensitivity, specificity and intensity of staining. In addition to providing an opportunity to use archived tissues, this new post-fixation method would dramatically reduce the use of formaldehyde during histopathology procedures, thus minimizing worker exposure to this dangerous carcinogenic substance.

• 出版日期2013