Biofilms: An ESEM evaluation of artifacts introduced during SEM preparation

Little, Brenda J.; Wagner, Patricia; Ray, Richard I.; Pope, Robert K.; Scheetz, Raymond W.

doi:10.1007/bf01576058

Cited by 152 publications

(94 citation statements)

References 18 publications

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“…4E). As noted by Little et al (236) for bacterial biofilms, solvent replacement of water from a biofilm removes matrix components. In addition, whether the SEM preparation is frozen or dehydrated after fixation, native architecture is lost.…”

Section: Methods For Measuring Biofilms May Be a Source Of Variabilitymentioning

confidence: 93%

Plasticity of Candida albicans Biofilms

Soll

Daniels

2016

Microbiol Mol Biol Rev

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SUMMARYCandida albicans, the most pervasive fungal pathogen that colonizes humans, forms biofilms that are architecturally complex. They consist of a basal yeast cell polylayer and an upper region of hyphae encapsulated in extracellular matrix. However, biofilms formedin vitrovary as a result of the different conditions employed in models, the methods used to assess biofilm formation, strain differences, and, in a most dramatic fashion, the configuration of the mating type locus (MTL). Therefore, integrating data from different studies can lead to problems of interpretation if such variability is not taken into account. Here we review the conditions and factors that cause biofilm variation, with the goal of engendering awareness that more attention must be paid to the strains employed, the methods used to assess biofilm development, every aspect of the model employed, and the configuration of theMTLlocus. We end by posing a set of questions that may be asked in comparing the results of different studies and developing protocols for new ones. This review should engender the notion that not all biofilms are created equal.

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Section: Methods For Measuring Biofilms May Be a Source Of Variabilitymentioning

confidence: 93%

Plasticity of Candida albicans Biofilms

Soll

Daniels

2016

Microbiol Mol Biol Rev

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“…High-resolution imaging such as scanning electron microscopy (SEM), or confocal laser scanning microscopy (CLSM) proved to be powerful tools for biofilm structure deciphering, but they require extensive sample preparation (SEM) or image analysis (CLSM) and are not suitable for routine use. An easier microscopic approach is the use of environmental scanning electron microscopy (ESEM), which allows the observation of microbial biofilms in their native state without fixation, dehydration or metal coating (Little et al, 1991;Alhede et al, 2012). The main advantages of biofilm ESEM imaging are, indeed, the visualization of the highly hydrated organic matrix surrounding the bacteria, and minimal processing of samples.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a non-submerged cultivation assay combined to ESEM imaging for analysis of biofilms formed by dairy-associated sporeforming bacteria

Malek¹

2016

Afr. J. Microbiol. Res.

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In the dairy industry, the biofilms formed by spore forming bacteria are not well characterized. Microscopic methods are crucial for the study of biofilm structural and architectural features. Here, a simple surface-associated non-submerged model combined to environmental scanning electron microscope (ESEM) imaging was used for the study of Bacillus cereus and Geobacillus spp. dairy biofilms. To evaluate the utility of this approach, non-submerged biofilms were compared to those developed in situ on stainless steel coupons introduced inside milk processing lines. Results reveal that both B. cereus and Geobacillus spp. are able to form specific biofilm characteristics on nonimmersed surfaces, notably an original dispersion style not previously described. Non-submerged biofilms in vitro are elaborate three-dimensional or extensive complex structures well resolved in ESEM and comparable to dairy biofilms in situ. The non-submerged surface-associated biofilm combined to ESEM imaging revealed a relevant model for the study of dairy biofilms.

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“…In contemporary research, several methods of biofilm characterisation using statistical techniques has been reported on 2D SEM images [42][43][44][45][46][47][48] . Yang et al 49 and Jackson et al 50 have studied several textural features of microscope images, e.g.…”

Section: Brief Survey Of the Existing Biofilm Image Analysis Techniquesmentioning

confidence: 99%

Monitoring of Wild Pseudomonas Biofilm Strain Conditions Using Statistical Characterization of Scanning Electron Microscopy Images

Sinha

Das

Tarafdar

et al. 2017

Ind. Eng. Chem. Res.

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Abstract:The present paper proposes a novel method of quantification of the variation in biofilm architecture, in correlation with the alteration of growth conditions that include, variations of substrate and conditioning layer. The polymeric biomaterial serving as substrates are widely used in implants and indwelling medical devices, while the plasma proteins serve as the conditioning layer. The present method uses descriptive statistics of FESEM images of biofilms obtained during a variety of growth conditions. We aim to explore here the texture and fractal analysis techniques, to identify the most discriminatory features which are capable of predicting the difference in biofilm growth conditions. We initially extract some statistical features of biofilm images on bare polymer surfaces, followed by those on the same substrates adsorbed with two different types of plasma proteins, viz. Bovine serum albumin (BSA) and Fibronectin (FN), for two different adsorption times. The present analysis has the potential to act as a futuristic technology for developing a computerized monitoring system in hospitals with automated image analysis and feature extraction, which may be used to predict the growth profile of an emerging biofilm on surgical implants or similar medical applications.

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Biofilms: An ESEM evaluation of artifacts introduced during SEM preparation

Cited by 152 publications

References 18 publications

Plasticity of Candida albicans Biofilms

Plasticity of Candida albicans Biofilms

Evaluation of a non-submerged cultivation assay combined to ESEM imaging for analysis of biofilms formed by dairy-associated sporeforming bacteria

Monitoring of Wild Pseudomonas Biofilm Strain Conditions Using Statistical Characterization of Scanning Electron Microscopy Images

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