Comparison of microscope techniques for the examination of biofilms

Surman, Susanne; Walker, Jimmy; Goddard, D. T.; Morton, L.H.G.; Keevil, C. W.; Weaver, Westbrook M.; Skinner, A.; Hanson, Kimberly L.; Caldwell, Douglas E.; Kurtz, J. B.

doi:10.1016/0167-7012(95)00085-2

Cited by 136 publications

(70 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The biofilms that have been imaged previously by AFM were all grown in fluid (2,4,55) or, in the case of P. putida, at an oil-water interface (20). Many of our AFM images of unsaturated P. putida are reminiscent of the published AFM images of P. putida in fluid (20,53), although the prior methods were quite different from ours.…”

Section: Discussionmentioning

confidence: 58%

Physical Morphology and Surface Properties of Unsaturated Pseudomonas putida Biofilms

et al. 2000

View full text Add to dashboard Cite

Unsaturated biofilms of Pseudomonas putida, i.e., biofilms grown in humid air, were analyzed by atomic force microscopy to determine surface morphology, roughness, and adhesion forces in the outer and basal cell layers of fresh and desiccated biofilms. Desiccated biofilms were equilibrated with a 75.5% relative humidity atmosphere, which is far below the relative humidity of 98 to 99% at which these biofilms were cultured. In sharp contrast to the effects of drying on biofilms grown in fluid, we observed that drying caused little change in morphology, roughness, or adhesion forces in these unsaturated biofilms. Surface roughness for moist and dry biofilms increased approximately linearly with increasing scan sizes. This indicated that the divides between bacteria contributed more to overall roughness than did extracellular polymeric substances (EPS) on individual bacteria. The EPS formed higher-order structures we termed mesostructures. These mesostructures are much larger than the discrete polymers of glycolipids and proteins that have been previously characterized on the outer surface of these gram-negative bacteria.

show abstract

Section: Discussionmentioning

confidence: 58%

Physical Morphology and Surface Properties of Unsaturated Pseudomonas putida Biofilms

et al. 2000

View full text Add to dashboard Cite

show abstract

“…It enables phylogenetic identification using 16s rRNAdirected oligonucleotide probes in addition to providing information on size, morphology and numbers of bacteria. The method has been further improved by CLSM, allowing optical sectioning, three-dimensional reconstruction and therefore exact localization and observation of the spatial distribution of microorganisms (Amann et al, 1996;MacNaughton et al, 1996;Manz et at., 1995;Surmann et al, 1996;Wilson, 1989;Wright & Schatten, 1991).…”

Section: Direct Visualization Of Bacteria By Fishmentioning

confidence: 99%

Fluorescence in situ hybridization shows spatial distribution of as yet uncultured treponemes in biopsies from digital dermatitis lesions

Moter¹,

et al. 1998

View full text Add to dashboard Cite

Fluorescence in situ hybridization (FISH) was performed on sections of plastic-embedded tissue using 16S rRNA-directed oligonucleotide probes to visualize uncultured treponemes in skin biopsies of cows with digital dermatitis. Plastic as embedding material allowed sectioning of hard and soft tissue with a defined thickness, avoiding the risk of dragging bacteria into the tissue while sectioning. Furthermore, it provided a good signal-to-noise ratio. Using this method the spatial distribution of three different bacterial phylotypes was visualized simultaneously within the tissue. Whereas debris covering the ulcers contained a mixture of different micro-organisms, a layering of certain treponemal phylotypes was observed deeper in the epidermis. Confocal laser scanning microscopy and subsequent three-dimensional reconstruction of series of optical sections confirmed that the treponemes migrated intercellularly around the cells, most of them directed towards the dermis. In situ hybridization on tissue embedded in plastic proved to be a useful method to study mixed bacterial infections since it combines excellent histological conservation of tissue with identification of bacterial species by simultaneous use of probes labelled with different fluorescent dyes. This technique may have implications for in situ detection, identification and localization of microorganisms in veterinary as well as in human medicine.

show abstract

“…For this reason, there is a shift in preparative procedures from dehydration of samples to techniques where partly dehydrated samples can be examined. Partly dehydrated samples may be employed for environmental SEM and for atomic force microscopy (Bremer et al 1992, Kasas et al 1994, Sutton et al 1994, Lavoie et al 1995, Surman et al 1996. Further, there has been the development and application of techniques allowing the observation of fully hydrated interfacial microbial consortia using confocal laser scanning microscopy (CLSM) (Caldwell et al 1992, Lawrence et al 1998a.…”

Section: Introductionmentioning

confidence: 99%

In situ cell and glycoconjugate distribution in river snow studied by confocal laser scanning microscopy

Neu¹

2000

Aquat. Microb. Ecol.

View full text Add to dashboard Cite

Aggregates from the river Elbe (Germany) were collected and examined by confocal laser scanning microscopy. The river snow was directly transferred into coverslip chambers and observed without any embedding and fixation procedure. River snow samples were examined in the reflection and fluorescence mode to record mineral content and autofluorescence signals. Spatial distribution of microb~al cells within the aggregates was shown with general nucleic acid specific stains (SYTO 9, SYTO 64). The polymeric matnx of the river snow was demonstrated by using a panel of fluorescently labelled lectins. Staining with FITC and TRITC labelled lectins and simultaneous dual channel unaging showed the distribution of different types of glycoconjugates. The binding pattern observed included bacterial cell surface labelling and cloud-type labelling of extracellular polymeric substances (EPS). In addition, dual staining combined with triple-channel recording was employed to clearly separate the signal in the red channel originating from TRITC-lectin or SYTO 64 nucleic acid stain from the far red autofluorescence originating from chlorophyll-containing organisms. With this approach the fully hydrated, living, 3-dimensional architecture of aquatic aggregates was investigated. Application of lectins demonstrated the heterogeneity of the EPS matrix in between the cellular constituents of the river snow community. The chemical heterogeneity of river snow may be significant for sorption and transport of nutrients and contaminants in lotlc aquatic environments. This in situ technique may be also useful to examine interfacial microbial consortia in other habitats. Finally, it is suggested to employ in situ lectin staining to probe for the glycoconjugate distribution at the polymer level similar to in situ hybridisation with rRNA targeted oligonucleotides at the cellular level.

show abstract

Comparison of microscope techniques for the examination of biofilms

Cited by 136 publications

References 12 publications

Physical Morphology and Surface Properties of Unsaturated Pseudomonas putida Biofilms

Physical Morphology and Surface Properties of Unsaturated Pseudomonas putida Biofilms

Fluorescence in situ hybridization shows spatial distribution of as yet uncultured treponemes in biopsies from digital dermatitis lesions

In situ cell and glycoconjugate distribution in river snow studied by confocal laser scanning microscopy

Contact Info

Product

Resources

About