Impact of fixatives on preservation of RNA in paraffin-embedded and laser capture microdissected human tissues

Authors: Park, In Suh1; Kim, Ye Ji1; Kim, Lucia1; Choi, Suk Jin1; Han, Jee Young1; Chu, Young Chae1; Kim, Han Kyum; Jin, Xianji1; Kim, Joon Mee1

Source: Basic and Applied Pathology, Volume 1, Number 2, June 2008 , pp. 72-76(5)

Publisher: Blackwell Publishing

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Abstract:

Background and aim: Laser capture microdissection is used to obtain pure cell populations, thus allowing the study of molecular pathological mechanisms from specific cell populations, yet RNA from tissues fixed in formalin (the most common fixative for analysis of gene expression) is usually degraded during microdissection and RNA extraction. In this study we investigate the best fixative for use in the preparation of human tissues for laser capture microdissection, and for the preservation of RNA molecules and histomorphologic structure. Methods: Here we examine Methacarn, Carnoy's solution, 4% paraformaldehyde and 10% neutral-buffered formalin. Forty-three sides of various human organs were compared to determine the tissue-fixing ability and RNA quality according to the fixatives. Results: Methacarn was the best suitable fixative for histomorphologic preservation as well as protection against RNA degradation after microdissection. However, Methacarn was not good for preservation of proteins. Conclusions: Methacarn is an excellent fixative for human tissue samples obtained by microdissection. It is an optimal fixative that is not only excellent for preservation of morphology, but also RNA extraction.

Keywords: formalin-fixed; laser capture microdissection; Methacarn; paraffin; RNA

Document Type: Original article

DOI: 10.1111/j.1755-9294.2008.00016.x

Affiliations: 1: Department of pathology, Inha University Hospital, Incheon

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