대한의생명과학회지 (Biomedical Science Letters)

  • 제29권1호
  • /
  • Pages.48-52
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  • 2023
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  • 1738-3226(pISSN)
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  • 2288-7415(eISSN)
대한의생명과학회 (The Korean Society for Biomedical Laboratory Sciences)
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Evaluating the Efficacy of a Formalin Alternative in Fixing Pathological Tissues for Histological and Molecular Diagnoses

  • Min-A Je (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Haneul Lee (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Heechul Park (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Dong Hyeok Kim (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Yeongdon Ju (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Jaewon Lim (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Sunghyun Kim (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan) ;
  • Jungho Kim (Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan)
  • 투고 : 2023.02.24
  • 심사 : 2023.03.29
  • 발행 : 2023.03.31
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초록

Formaldehyde use is associated with serious health risks, which can affect medical personnel and technicians. Therefore, we investigated the efficacy of an alternative fixative, with respect to two types of formalin fixatives, by hematoxylin and eosin (H&E) staining, periodic acid Schiff (PAS) staining, immunohistochemical (IHC) staining, and RNA extraction. For H&E staining, the circular nucleus was stained dark blue by the basic dye hematoxylin and the cytoplasm was stained red by the acid dye eosin in all three fixative samples. No difference was found in the Duksan General Science (DGS), Sigma-Aldrich, and Core-Fix fixative samples (Corebiotech) used to fix kidney tissue, after PAS staining. IHC staining showed that CD4 was significantly increased in the lippolysaccharide (LPS)-treated group compared to the control group (vehicle), confirming the changes in specific molecules. The quantity and quality of RNA from tissues fixed in the three types of fixatives were evaluated. The average concentration of RNA was 106 ng/µL and average purity at A 260/280 ratio was 1.7~2.0, regardless of fixative used. For quality of protein, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein was confirmed by Western blotting. In conclusion, Core-Fix can be used as a fixative for pathological tissues, in histological and molecular diagnoses.

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참고문헌

  1. ACGIH. "TLVs® and BEIs®: Threshold Limit Values for Chemical Substances and Physical Agents and Biological Exposure Indices Available from 2013: URL: http://www.acgih.org/science/tlv-bei-guidelines/
  2. Alexandersson R, Hedenstierna G. "Pulmonary function in wood workers exposed to formaldehyde: a prospective study". Arch Environ Health. 1989. 44: 5-11. https://doi.org/10.1080/00039896.1989.9935865
  3. Bass BP, Engel KB, Greytak SR, Moore HM. "A review of pre-analytical factors affecting molecular, protein, and morphological analysis of formalin-fixed, paraffin-embedded (FFPE) tissue: how well do you know your FFPE specimen?" Arch Pathol Lab Med. 2014. 138: 1520-1530. https://doi.org/10.5858/arpa.2013-0691-RA
  4. Becker KF, Schott C, Hipp S, Metzger V, Porschewski P, Beck R, Nahrig J, Becker I, Hofler H. "Quantitative protein analysis from formalin-fixed tissues: implications for translational clinical research and nanoscale molecular diagnosis". J Pathol. 2007. 211: 370-378. https://doi.org/10.1002/path.2107
  5. Bronsert P, Weisser J, Biniossek ML, Kuehs M, Mayer B, Drendel V, Timme S, Shahinian H, Kusters S, Wellner UF, Lassmann S, Werner M, Schilling O. "Impact of routinely employed procedures for tissue processing on the proteomic analysis of formalin-fixed paraffin-embedded tissue". Proteomics Clin Appl. 2014. 8: 796-804. https://doi.org/10.1002/prca.201300082
  6. Buesa RJ. "Histology without formalin?" Ann Diagn Pathol. 2008. 12: 387-396. https://doi.org/10.1016/j.anndiagpath.2008.07.004
  7. Costa S, Coelho P, Costa C, Silva S, Mayan O, Santos LS, Gaspar J, Teixeira JP. "Genotoxic damage in pathology anatomy laboratory workers exposed to formaldehyde". Toxicology. 2008. 252: 40-48. https://doi.org/10.1016/j.tox.2008.07.056
  8. Hauptmann M, Lubin JH, Stewart PA, Hayes RB, Blair A. "Mortality from solid cancers among workers in formaldehyde industries". Am J Epidemiol. 2004. 159: 1117-1130. https://doi.org/10.1093/aje/kwh174
  9. IARC. "Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans Available from 2013: URL: http://monographs.iarc.fr/ENG/Classification/index.php".
  10. Kothmaier H, Rohrer D, Stacher E, Quehenberger F, Becker KF, Popper HH. "Comparison of formalin-free tissue fixatives: a proteomic study testing their application for routine pathology and research". Arch Pathol Lab Med. 2011. 135: 744-752. https://doi.org/10.5858/2009-0676-OA.1
  11. Ludyga N, Grunwald B, Azimzadeh O, Englert S, Hofler H, Tapio S, Aubele M. "Nucleic acids from long-term preserved FFPE tissues are suitable for downstream analyses". Virchows Arch. 2012. 460: 131-140. https://doi.org/10.1007/s00428-011-1184-9
  12. Nirmalan NJ, Harnden P, Selby PJ, Banks RE. "Development and validation of a novel protein extraction methodology for quantitation of protein expression in formalin-fixed paraffin-embedded tissues using western blotting". J Pathol. 2009. 217: 497-506. https://doi.org/10.1002/path.2504
  13. Puchtler H, Meloan SN. "On the chemistry of formaldehyde fixation and its effects on immunohistochemical reactions". Histochemistry. 1985. 82: 201-204. https://doi.org/10.1007/BF00501395
  14. Qin Y, Jiang W, Li A, Gao M, Liu H, Gao Y, Tian X, Gong G. "The Combination of Paraformaldehyde and Glutaraldehyde Is a Potential Fixative for Mitochondria". Biomolecules. 2021. 11.
  15. Zeng F, Yang W, Huang J, Chen Y, Chen Y. "Determination of the lowest concentrations of aldehyde fixatives for completely fixing various cellular structures by real-time imaging and quantification". Histochem Cell Biol. 2013. 139: 735-749. https://doi.org/10.1007/s00418-012-1058-5