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Göteborgs universitets publikationer

Influence of fixation method on the accuracy of assessment of mRNA in cells with the In Situ hybridization technique

Författare och institution:
Lena Larsson (Institutionen för odontologi); Tord Berglundh (Institutionen för odontologi)
Publicerad i:
Journal of Histotechnology, 31 ( 2 ) s. 45-50
Artikel, refereegranskad vetenskaplig
Sammanfattning (abstract):
Objective: The aim of this study was to evaluate the reproducibility and accuracy of mRNA detection using two methods for tissue preservation. Material and methods: Soft tissue biopsies from inflamed gingival tissues were collected from subjects with generalized or localized forms of severe periodontitis or periimplantitis, i.e. inflammatory destructive disease in the tissues surrounding teeth and dental implants. The In Situ hybridization technique was used to compare detection of mRNA in paraformaldehyde-fixed tissue with frozen tissue. The histological quantitative assessments were performed using a point counting procedure to determine the percentage of positive cell markers within the defined infiltrated connective tissue (ICT). In order to estimate the error of measurements double assessments were performed with a two months interval. Results: It was demonstrated that assessments of cell proportions in sections from unfixed, frozen tissue resulted in larger standard deviations than assessments made in sections prepared from PFA-fixed tissue samples. A significantly larger density of IL-10 positive cells was found in the sections prepared from PFA fixed tissue than in sections from frozen tissue samples. It is suggested that paraformaldehyde fixation of tissues is superior to the use of frozen tissue in the preparation of tissue samples for detection of mRNA detection using In Situ Hybridization. Conclusion: The variability between the two preservation techniques presented in the current study has to be considered in the planning of studies evaluating differences between densities of cell markers in tissue samples prepared for ISH.
Ämne (baseras på Högskoleverkets indelning av forskningsämnen):
Klinisk medicin ->
Odontologi ->
histology, human, ISH, mRNA, probe
Postens nummer:
Posten skapad:
2008-08-04 09:15
Posten ändrad:
2008-08-05 13:43

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