scholarly journals High Rates of Conjugation in Bacterial Biofilms as Determined by Quantitative In Situ Analysis

1999 ◽  
Vol 65 (8) ◽  
pp. 3710-3713 ◽  
Author(s):  
Martina Hausner ◽  
Stefan Wuertz

ABSTRACT Quantitative in situ determination of conjugative gene transfer in defined bacterial biofilms using automated confocal laser scanning microscopy followed by three-dimensional analysis of cellular biovolumes revealed conjugation rates 1,000-fold higher than those determined by classical plating techniques. Conjugation events were not affected by nutrient concentration alone but were influenced by time and biofilm structure.

2004 ◽  
Vol 49 (11-12) ◽  
pp. 177-185 ◽  
Author(s):  
J.B. Xavier ◽  
C. Picioreanu ◽  
M.C.M. van Loosdrecht

The mathematical modeling of spatial biofilm formation that provides the capability to predict biofilm structure from first principles has been in development for the past six years. However, a direct and quantitative link between model predictions and the experimentally observed structure formation still remains to be established. This work assesses the capability of a state-of-the-art technique for three-dimensional (3D) modeling of biofilm structure, individual based modeling (IbM), to quantitatively describe the early development of a multispecies denitrifying biofilm. Model evaluation was carried out by comparison of predicted structure with that observed from two experimental datasets using confocal laser scanning microscopy (CLSM) monitoring of biofilm development in laboratory flowcells. Experimental conditions provided biofilm growth without substrate limitation, which was confirmed from substrate profiles computed by the model. 3D structures were compared quantitatively using a set of morphological parameters including the biovolume, filled-space profiles, substratum coverage, average thickness and normalized roughness. In spite of the different morphologies detectable in the two independent short-term experiments analyzed here, the model was capable of accurate fitting data from both experiments. Prediction of structure formation was precise, as expressed by the set of morphology parameters used.


RSC Advances ◽  
2016 ◽  
Vol 6 (103) ◽  
pp. 101235-101241 ◽  
Author(s):  
Pengcheng Yan ◽  
Yeon Joo Jeong ◽  
Mohammad F. Islam ◽  
P. Chris Pistorius

Determination of graphene formation and growth using direct,in situimaging with high-temperature laser scanning confocal microscope.


2007 ◽  
Vol 55 (8-9) ◽  
pp. 207-210 ◽  
Author(s):  
D. Spettmann ◽  
S. Eppmann ◽  
H.-C. Flemming ◽  
J. Wingender

Fouling is a major problem in membrane processes of water treatment. It can be caused by the deposition of inorganic and organic particulate material, and of microbial cells which may subsequently form biofilms. In practice, usually more than one foulant participates in the formation of membrane deposits. Knowledge of the composition of fouling layers is important for the development of appropriate countermeasures. For this purpose, an experimental system was established for the generation and microscopic visualisation of mixed deposits, using fluorescently labelled model foulants: (i) drinking-water bacteria stained with nucleic acid-specific dyes (biofouling), (ii) synthetic clay mineral laponite stained with rhodamine 6G (inorganic particle fouling), and (iii) fluorescently labelled polystyrene microspheres (organic particle fouling). Polycarbonate and polyethersulfone membranes were challenged with these foulants by dead-end filtration. On the basis of different fluorescent labels, the single foulants in these mixed deposits could be visualised separately by confocal laser scanning microscopy which, in combination with image analysis, allowed the generation of three-dimensional views of the complete deposits. This method offers the possibility for the estimation of quantitative surface coverage by foulants and for the determination of the efficacy of cleaning measures with respect to the removal of different foulants.


Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.


Materials ◽  
2021 ◽  
Vol 14 (8) ◽  
pp. 1821
Author(s):  
Ting He ◽  
Wei Shi ◽  
Song Xiang ◽  
Chaowen Huang ◽  
Ronald G. Ballinger

The influence of AlFeSi and Mg2Si phases on corrosion behaviour of the cast 6061 aluminium alloy was investigated. Scanning Kelvin probe force microscopy (SKPFM), electron probe microanalysis (EPMA), and in situ observations by confocal laser scanning microscopy (CLSM) were used. It was found that Mg2Si phases were anodic relative to the matrix and dissolved preferentially without significantly affecting corrosion propagation. The AlFeSi phases’ influence on 6061 aluminium alloy local corrosion was greater than that of the Mg2Si phases. The corroded region width reached five times that of the AlFeSi phase, and the accelerating effect was terminated as the AlFeSi dissolved.


Life ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 301
Author(s):  
Kathrin Malfeld ◽  
Nina Armbrecht ◽  
Holger A. Volk ◽  
Thomas Lenarz ◽  
Verena Scheper

In recent years sensorineural hearing loss was found to affect not exclusively, nor at first, the sensory cells of the inner ear. The sensory cells’ synapses and subsequent neurites are initially damaged. Auditory synaptopathies also play an important role in cochlear implant (CI) care, as they can lead to a loss of physiological hearing in patients with residual hearing. These auditory synaptopathies and in general the cascades of hearing pathologies have been in the focus of research in recent years with the aim to develop more targeted and individually tailored therapeutics. In the current study, a method to examine implanted inner ears of guinea pigs was developed to examine the synapse level. For this purpose, the cochlea is made transparent and scanned with the implant in situ using confocal laser scanning microscopy. Three different preparation methods were compared to enable both an overview image of the cochlea for assessing the CI position and images of the synapses on the same specimen. The best results were achieved by dissection of the bony capsule of the cochlea.


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