Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1099/00221287-147-2-299
Title (Primary) Assessment of lectin-binding analysis for in situ detection of glycoconjugates in biofilm systems
Author Neu, T.R.; Swerhone, G.D.W.; Lawrence, J.R.
Source Titel Microbiology-SGM
Year 2001
Department FLOEK; GM
Volume 147
Issue 2
Page From 299
Page To 313
Language englisch
Keywords bio®lms, lectins, extracellular polymeric substances, glycoconjugates, confocal laser scanning microscopy
Abstract An assessment of lectin-binding analysis for the characterization of extracellular glycoconjugates as part of the extracellular polymeric substances in environmental microbial communities was performed using fully hydrated river biofilms. The applicability of the method was evaluated for single, dual and triple staining with a panel of fluor-conjugated lectins. It was shown that lectin-binding analysis was able to stain glycoconjugates within biofilm communities. Lectin staining also demonstrated spatial heterogeneity within
the biofilm matrix. Furthermore, the application of two or even three lectins was possible if suitable combinations were selected. The lectin-binding analysis can be combined with general nucleic acid stains to collect both nucleic acid and glycoconjugate signals. The effects of incubation time, lectin concentration, fluor labelling, carbohydrate inhibition, order of addition and lectin interactions were studied. An incubation time of 20 min was found to be sufficient for completion of lectin binding. It was not possible to ascertain
saturating concentration for individual lectins, therefore a standard concentration was used for the assay. Carbohydrate inhibition tests indicated that fluorescein isothiocyanate (FITC)-conjugated lectins had more specific binding characteristics than tetramethyl rhodamine isothiocyanate (TRITC)- or cyanine dye (CY5)-labelled lectins. The order of addition and the nature of the fluor conjugate were also found to influence the binding pattern of the lectins. Therefore the selection of a panel of lectins for investigating the EPS matrix
must be based on a full evaluation of their behaviour in the biofilm system to be studied. Despite this necessity, lectin-binding analysis represents a valuable tool to examine the glycoconjugate distribution in fully hydrated biofilms. Thereby, chemical heterogeneities within extracellular biofilm locations can be identified in order to examine the role (e.g. sorption properties, microenvironments, cell–extracellular polymeric subtance interactions) of the extracellular polymeric substances in environmental biofilm systems.
Persistent UFZ Identifier
Neu, T.R., Swerhone, G.D.W., Lawrence, J.R. (2001):
Assessment of lectin-binding analysis for in situ detection of glycoconjugates in biofilm systems
Microbiology-(UK) 147 (2), 299 - 313 10.1099/00221287-147-2-299