Optimization of fixation methods for observation of bacterial cell morphology and surface ultrastructures by atomic force microscopy

Chao, Yuanqing; Zhang, Tong

doi:10.1007/s00253-011-3551-5

Cited by 165 publications

(115 citation statements)

References 37 publications

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“…The number of microcolonies that were attached on the reactor slides was calculated directly on the slides of the reactor by fluorescence microscopy using the 100× magnification after the samples were fixed with 0.5 mL of 4% paraformaldehyde [58]. Also, the number of aggregates was determined directly by visual observation on the reactor slides.…”

Section: Flow Conditionsmentioning

confidence: 99%

A Keystone Methylobacterium Strain in Biofilm Formation in Drinking Water

Tsagkari

Keating

Couto

et al. 2017

Water

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Abstract:The structure of biofilms in drinking water systems is influenced by the interplay between biological and physical processes. Bacterial aggregates in bulk fluid are important in seeding biofilm formation on surfaces. In simple pure and co-cultures, certain bacteria, including Methylobacterium, are implicated in the formation of aggregates. However, it is unclear whether they help to form aggregates in complex mixed bacterial communities. Furthermore, different flow regimes could affect the formation and destination of aggregates. In this study, real drinking water mixed microbial communities were inoculated with the Methylobacterium strain DSM 18358. The propensity of Methylobacterium to promote aggregation was monitored under both stagnant and flow conditions. Under stagnant conditions, Methylobacterium enhanced bacterial aggregation even when it was inoculated in drinking water at 1% relative abundance. Laminar and turbulent flows were developed in a rotating annular reactor. Methylobacterium was found to promote a higher degree of aggregation in turbulent than laminar flow. Finally, fluorescence in situ hybridisation images revealed that Methylobacterium aggregates had distinct spatial structures under the different flow conditions. Overall, Methylobacterium was found to be a key strain in the formation of aggregates in bulk water and subsequently in the formation of biofilms on surfaces.

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Section: Flow Conditionsmentioning

confidence: 99%

A Keystone Methylobacterium Strain in Biofilm Formation in Drinking Water

Tsagkari

Keating

Couto

et al. 2017

Water

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“…Samples were fixed with 2.5% glutaraldehyde in PBS (glutaraldehyde from Arcos Organics). 30 Fixed bacteria were examined using tapping-mode AFM (NT-MDT, NTEGRA) with a 10 nm radius of curvature tip (ACL-10, Applied NanoStructures, Mountain View, CA), or sputter coated with iridium and imaged under high-vacuum SEM (FEI Quanta 3D). Lengths of individual bacterial cells and their angular orientations with respect to the pillar arrays were measured from the SEM images using IMAGEJ software.…”

Section: E Bacterial Morphologymentioning

confidence: 99%

Nanopatterned polymer surfaces with bactericidal properties

Dickson¹,

Liang²,

Rodríguez³

et al. 2015

Biointerphases

233

240

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Bacteria that adhere to the surfaces of implanted medical devices can cause catastrophic infection. Since chemical modifications of materials' surfaces have poor long-term performance in preventing bacterial buildup, approaches using bactericidal physical surface topography have been investigated. The authors used Nanoimprint Lithography was used to fabricate a library of biomimetic nanopillars on the surfaces of poly(methyl methacrylate) (PMMA) films. After incubation of Escherichia coli (E. coli) on the structured PMMA surfaces, pillared surfaces were found to have lower densities of adherent cells compared to flat films (67%-91% of densities on flat films). Moreover, of the E. coli that did adhere a greater fraction of them were dead on pillared surfaces (16%-141% higher dead fraction than on flat films). Smaller more closely spaced nanopillars had better performance. The smallest most closely spaced nanopillars were found to reduce the bacterial load in contaminated aqueous suspensions by 50% over a 24-h period compared to flat controls. Through quantitative analysis of cell orientation data, it was determined that the minimum threshold for optimal nanopillar spacing is between 130 and 380 nm. Measurements of bacterial cell length indicate that nanopillars adversely affect E. coli morphology, eliciting a filamentous response. Taken together, this work shows that imprinted polymer nanostructures with precisely defined geometries can kill bacteria without any chemical modifications. These results effectively translate bactericidal nanopillar topographies to PMMA, an important polymer used for medical devices.

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“…In general, the fixation procedures, including chemical or physical methods, would change cell morphologies and mechanical properties. Aldehyde-based fixatives are useful for conserving cell or bacterial morphologies [60,61]. Nonetheless, it is very possible that different samples require different notes: This figure is adopted from Ref.…”

Section: Cell Membrane Responses To Chemical Stimulationsmentioning

confidence: 99%

Mechanoprofiling on membranes of living cells with atomic force microscopy and optical nano-profilometry

Lee

Wang

Lee

2017

Advances in Physics: X

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Membrane topography of living cells has been considered as an effective parameter that reflects cellular statuses. With the improvements in spatial and temporal resolutions of various measurement techniques, the changes of membrane topography in response to various external stimulations in the culture environments can be accurately recorded. Membrane roughness is a useful parameter to evaluate the changes in membrane topography. At present, atomic force microscopy (AFM) is the most common technique to measure the membrane topography and roughness of living cells. On the other hand, with the non-contact profiling capability, many optical techniques are also employed in this field of research. This review briefly introduces the evolution and applications of AFM on measuring membrane roughness. Meanwhile, quantifying the cell membrane topography and roughness with a non-scanning, non-contact, and label-free optical technique, non-interferometric wide-field optical profilometry (NIWOP), is also presented.

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Optimization of fixation methods for observation of bacterial cell morphology and surface ultrastructures by atomic force microscopy

Cited by 165 publications

References 37 publications

A Keystone Methylobacterium Strain in Biofilm Formation in Drinking Water

A Keystone Methylobacterium Strain in Biofilm Formation in Drinking Water

Nanopatterned polymer surfaces with bactericidal properties

Mechanoprofiling on membranes of living cells with atomic force microscopy and optical nano-profilometry

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