에탄올 고정액의 조직병리학적, 분자생물학적 분석 유효성 평가
Validity Assessment of Ethanol for Fixatives of Histopathological and Molecular Biological Analysis
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초록
As the global importance of aquaculture grows, academic interest in aquatic diseases is also increasing. Diagnosis of aquatic diseases is made through expert diagnosis, and it is recommended to use histopathological and molecular methods in parallel. Neutral buffered formalin (NBF) is used as a conventional tissue fixative. However, its toxicity in standard laboratories complicates handling and precludes molecular biological analysis. As a result, there has been ongoing scholarly effort toward alcohol-based fixatives that can accommodate both histopathological and molecular diagnostic modalities. In this Study, we closely examined the efficacy of ethanol fixatives at concentrations below 70%, rather than the traditionally explored >70% concentrations, assessing the resulting morphological changes and nucleic acid concentrations. Subsequently, an evaluation of the viability of ethanol fix- atives for histopathological and molecular diagnostic applications was performed, depending on the concentration and duration of exposure. Studies have shown that the use of 15% ethanol fixative dramatically reduces nucleic acid yield and induces severe morphological changes from the start. In contrast, application of 40% ethanol fixative induced moderate morphological changes and main- tained nucleic acid yield for up to 1 day. These fixatives also produce moderate morphological changes that persist for up to 3 days, although noticeable morphological changes occur sporadically. This demonstrates the feasibility of using 40% ethanol fixative for short-term fixation periods not exceeding 3 days. Nevertheless, further investigation is needed to elucidate subtle temporal dynamics and temper- ature effects, thus further studies are needed to confirm the feasibility of the 40% ethanol fixative.
Abstract
As the global importance of aquaculture grows, academic interest in aquatic diseases is also increasing. Diagnosis of aquatic diseases is made through expert diagnosis, and it is recommended to use histopathological and molecular methods in parallel. Neutral buffered formalin (NBF) is used as a conventional tissue fixative. However, its toxicity in standard laboratories complicates handling and precludes molecular biological analysis. As a result, there has been ongoing scholarly effort toward alcohol-based fixatives that can accommodate both histopathological and molecular diagnostic modalities. In this Study, we closely examined the efficacy of ethanol fixatives at concentrations below 70%, rather than the traditionally explored >70% concentrations, assessing the resulting morphological changes and nucleic acid concentrations. Subsequently, an evaluation of the viability of ethanol fix- atives for histopathological and molecular diagnostic applications was performed, depending on the concentration and duration of exposure. Studies have shown that the use of 15% ethanol fixative dramatically reduces nucleic acid yield and induces severe morphological changes from the start. In contrast, application of 40% ethanol fixative induced moderate morphological changes and main- tained nucleic acid yield for up to 1 day. These fixatives also produce moderate morphological changes that persist for up to 3 days, although noticeable morphological changes occur sporadically. This demonstrates the feasibility of using 40% ethanol fixative for short-term fixation periods not exceeding 3 days. Nevertheless, further investigation is needed to elucidate subtle temporal dynamics and temper- ature effects, thus further studies are needed to confirm the feasibility of the 40% ethanol fixative.
참고문헌 (47)
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