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|Title: ||A SHORTENED RNA-FISH PROTOCOL FOR ANALYZING ARTWORKS’ MICROCOLONIZERS|
|Authors: ||González-Pérez, Marina|
de Oliveira Baptista, Sara Margarida
Caldeira, Ana Teresa
|Keywords: ||Fluorescence In Situ Hybridization|
|Issue Date: ||4-Jan-2017|
|Publisher: ||YILDIZ TECHNICAL UNIVERSITY’S TECHNOPARK COMPANY OF PROMECH TEKNOLOJİ VE BİLİŞİM SİSTEMLERİ SANAYİ LTD. ŞTİ.|
|Citation: ||González-Pérez, M; de Oliveira Baptista, S.M.; Pereira, A; Candeias, A.; Caldeira, A.T.; A shortened RNA-FISH protocol for analyzing artworks' microcolonizers.Istanbul International Conference On Progress In Applied Science 2017 (ICPAS 2017), p. 136-140. ISBN 978-605-9546-02-7.
Istambul, Turkey, January 4-6, 2017.|
|Abstract: ||Microorganisms are involved in the deterioration of Cultural Heritage. Thus, there is a need to enhance the techniques used for their detection and identification.
RNA Fluorescent In Situ Hybridization (RNA-FISH) has been successfully applied for phylogenetic identification of the viable components of the microbial communities colonizing artworks both in situ and ex situ. Until recently, it was time-consuming, taking not less than 6 h for the analysis. We have developed an RNA-FISH in suspension protocol that allowed ex situ analysis of microorganisms involved in artworks’ biodeterioration in 5 h. In this work, three modified protocols, involving microwave heating, were evaluated for further shortening two of the four main critical steps in RNA-FISH: hybridization and washing.
The original and modified protocols were applied in cellular suspensions of bacteria and yeast isolates. The results obtained were evaluated and compared in terms of detectability and specificity of the signals detected by epifluorescence microscopy.
One of the methods tested showed good and specific FISH signals for all the microorganisms selected and did not produce signals evidencing non-specific or fixation-induced fluorescence.
This 3 h protocol allows a remarkable reduction of the time usually required for performing RNA-FISH analysis in Cultural Heritage samples. Thus, a rapid alternative for analyzing yeast and bacteria cells colonizing artworks’ surfaces by RNA-FISH is presented in this work.|
|Appears in Collections:||HERCULES - Artigos em Livros de Actas/Proceedings|
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