Scanning Electron Microscopy of Bacterial Colonies

Afrikian, E. G.; Julian, Grant St.; Bulla, Lee A.

doi:10.1128/am.26.6.934-937.1973

Cited by 8 publications

(4 citation statements)

References 9 publications

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“…However, most preparative procedures only applied glutaraldehyde fixation for one to two hours to get well-preserved bacterium morphology (Allan-Wojtas et al, 2008;Forge et al, 1992;Fratesi et al, 2004). In the case of formaldehyde, 24 hours of fixing is sufficient in the usual process (Williams, Bloebaum, 2010;Afrikian et al, 1973). The regularly used fixative 10% formalin is a low-cost, widely accessible fixative that does not induce significant shrinkage or cellular structural deformation (Thavarajah et al, 2012).…”

Section: Resultsmentioning

confidence: 99%

“…Cell-substrate interaction is important for less cell removal from the surface while treating with chemicals. Dialysis tubing was reported to be an excellent material since it supports bacterial growth, readily available, easily handled, sterilizable, withstand chemical fixation, drying, vacuum pressure, and remain undistorted under electron bombardment in the scanning microscope (Afrikian et al, 1973). Dialysis tubing was used to study the growth of an epiphytic diatom and bacterial community on its surface and the SEM results revealed clear bacteria morphology and biofilms (Vargo et al, 1975).…”

Section: Effect Of Substrate Materials On Sem Images Of P Aeruginosamentioning

confidence: 99%

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Sample preparative procedure for Pseudomonas aeruginosa observation under scanning electron microscope

Nguyen

Pham

Huynh

et al. 2022

Vietnam J. Biotechnol.

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Scanning electron microscope (SEM) is a popular tool used for observing bacteria surface and morphology. Using Pseudomonas aeruginosa as a model, this work aimed to show a SEM preparative procedure that is simple and economical but does not result in considerable data loss. This was accomplished via testing fixing ability of 10% formalin versus 2.5% glutaraldehyde, efficiency of air drying versus t-butyl alcohol drying method. Following that, polypropylene, dialysis tubing and agar were also assessed for their ability to serve as a supporting material for cell adhesion in preparing sample for SEM. Consequently, obtained data showed that the procedure using 24-hour 10% formalin fixation and t-butyl alcohol drying preserved well bacterial morphology. With this procedure, little cell or membrane damage was seen while extracellular structures were clearly observed. Furthermore, when this procedure was applied with different types of substrates including polypropylene, dialysis tubing, and agar, it showed that sample fixed on polypropylene maintained well extracellular structures meanwhile sample fixed on agar presented well bacterial morphology. In conclusion, our data suggested that coating samples on polypropylene, followed by 24-hour 10% formalin fixation and t-butyl alcohol drying was appropriate for observing bacteria under SEM.

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Section: Resultsmentioning

confidence: 99%

Section: Effect Of Substrate Materials On Sem Images Of P Aeruginosamentioning

confidence: 99%

Sample preparative procedure for Pseudomonas aeruginosa observation under scanning electron microscope

Nguyen

Pham

Huynh

et al. 2022

Vietnam J. Biotechnol.

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“…Isolates E. coli( 11161644), Klebsiella pneumoniae (11176977), Pseudomonas aeruginosa(11093627) as Gve bacteria, and Staphylococcus aureus (MRSA) (11182821) as G + ve bacteria were prepared to be examined using SEM, according to a method of Afrikian, et al, (1973) as follows: 1) Inoculate NB medium tube with bacteria isolated from bloodstream as a control. 2) Incubate inoculated tubes for 18 hours at 37C.…”

Section: Scanning Electron Microscope (Sem) Studiesmentioning

confidence: 99%

Synthesis of Rod/Spherical-Gold Nanoparticles (AuNPs) to Overcome Multidrug-Resistant Bacteremia

Shoeib

Hindi²,

Alsayed

2021

Alexandria Science Exchange Journal

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Multidrug-resistant bacteremia 15 Gve and 1 G + ve isolates were collected from inpatients and outpatients departments at Security Forces Hospital in Riyadh, Saudi Arabia. In studying the multidrug-resistance to antibiotics, it was recorded that the Gve isolates of E. coli (11093623) & (11161644) were resistance to 19 out of 20 tested antibiotics except antibiotic(CN), while the G + ve isolate of MRSA was resistance to 9 out of 18 antibiotics, and it was sensitive to antibiotic (OX and CZ).The Gold Nanoparticles (AuNPs) was examined to discourage the growth of the multidrug-resistance isolates. Rod-AuNPs were the best to discourage the growth of tested bacteria, followed by the Spherical-AuNPs. Swabs were taken from inhibition zone resulting from the inhibitory effect of AuNPs, towards the tested bacteria and re-inoculated on the Nutrient Agar medium, to determine either the bacteriostatic/bactericidal effect for different NPs. The study proved that Rod-AuNPs gave bacteriostatic effect on Pseudomonas aeruginosa (11093627) & (11132667), Enterobacter cloacae (11159053) and Citrobacter freundii(11176991)&(11182565), and gave a bactericidal effect to the rest of the tested isolates. The Spherical-AuNPs gave a bacteriostatic impact of all tested isolates. Electromicroscopic studies disclosed the effect of AuNPs, either rods or spherical-shaped, on the external shape of some of the pathogenic tested bacteria e.g. E. coli, Klebsiella pneumoniae and P. aeruginosa of Gve bacteria, MRSA of G + ve bacteria. From results obtained, it can be concluded that the nanostructures and morphologies of AuNPs may offer new possibilities for promising therapeutic applications of multidrug-resistance bacteremia.

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“…Field-of-view (FOV) dimensions accessible by SEM enable capturing statistically significant numbers of resolved individual cells within a single image [Figures S1–S2 in the Supporting Information (SI)]. , The high spatial resolution and depth-of-field of SEM also enable simultaneous observation of μm-scale objects, such as bacteria, and NPs resolved individually or in aggregates (Figure ).…”

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confidence: 99%

Analytical Protocols for Separation and Electron Microscopy of Nanoparticles Interacting with Bacterial Cells

et al. 2015

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An important step toward understanding interactions between nanoparticles (NPs) and bacteria is the ability to directly observe NPs interacting with bacterial cells. NP-bacteria mixtures typical in nanomedicine, however, are not yet amendable for direct imaging in solution. Instead, evidence of NP-cell interactions must be preserved in derivative (usually dried) samples to be subsequently revealed in high-resolution images, for example, via scanning electron microscopy (SEM). Here, this concept is realized for a mixed suspension of model NPs and Staphylococcus aureus bacteria. First, protocols for analyzing the relative colloidal stabilities of NPs and bacteria are developed and validated based on systematic centrifugation and comparison of colony forming unit (CFU) counting and optical density (OD) measurements. Rate-dependence of centrifugation efficiency for each component suggests differential sedimentation at a specific predicted rate as an effective method for removing free NPs after co-incubation; the remaining fraction comprises bacteria with any associated NPs and can be examined, for example, by SEM, for evidence of NP-bacteria interactions. These analytical protocols, validated by systematic control experiments and high-resolution SEM imaging, should be generally applicable for investigating NP-bacteria interactions.

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Scanning Electron Microscopy of Bacterial Colonies

Cited by 8 publications

References 9 publications

Sample preparative procedure for Pseudomonas aeruginosa observation under scanning electron microscope

Sample preparative procedure for Pseudomonas aeruginosa observation under scanning electron microscope

Synthesis of Rod/Spherical-Gold Nanoparticles (AuNPs) to Overcome Multidrug-Resistant Bacteremia

Analytical Protocols for Separation and Electron Microscopy of Nanoparticles Interacting with Bacterial Cells

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