pH-FISH: coupled microscale analysis of microbial identity and acid–base metabolism in complex biofilm samples

Yumi Chokyu Del Rey (Korresp. Autor*in), Katharina Kitzinger, Marie Braad Lund, Andreas Schramm, Rikke Louise Meyer, Michael Wagner, Sebastian Schlafer (Korresp. Autor*in)

Veröffentlichungen: Beitrag in FachzeitschriftArtikelPeer Reviewed

Abstract

Background: Correlative structural and chemical imaging of biofilms allows for the combined analysis of microbial identity and metabolism at the microscale. Here, we developed pH-FISH, a method that combines pH ratiometry with fluorescence in situ hybridization (FISH) in structurally intact biofilms for the coupled investigation of microbial acid metabolism and biofilm composition. Careful biofilm handling and modified sample preparation procedures for FISH allowed preservation of the three-dimensional biofilm structure throughout all processing and imaging steps. We then employed pH-FISH to investigate the relationship between local biofilm pH and the distribution of acid-producing (streptococci) and acid-consuming (Veillonella spp.) bacteria in dental biofilms from healthy subjects and caries-active patients. Results: The relative abundance of streptococci correlated with low biofilm pH at the field-of-view level, while the opposite trend was observed for Veillonella spp. These results suggest that clusters of streptococci contribute to the formation of acidic pockets inside dental biofilms, whereas Veillonella spp. may have a protective role against biofilm acidification. Conclusions: pH-FISH combines microscale mapping of biofilm pH in real time with structural imaging of the local microbial architecture, and is a powerful method to explore the interplay between biofilm composition and metabolism in complex biological systems.

OriginalspracheEnglisch
Aufsatznummer266
Seitenumfang11
FachzeitschriftMicrobiome
Jahrgang12
Ausgabenummer1
DOIs
PublikationsstatusVeröffentlicht - 20 Dez. 2024

ÖFOS 2012

  • 106022 Mikrobiologie
  • 105205 Klimawandel

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