Comparison of Alcian blue and ruthenium red effects on preservation of outer envelope ultrastructure in methanotrophic bacteria

Fassel, Theresa A.; Schaller, Marilyn; Remsen, Charles C.

doi:10.1002/jemt.1070200109

Cited by 14 publications

(12 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Likewise, the dramatic visualization of glycocalyx in E. faecalis biofilms seen with alcian blue is quite distinct from that seen with safranin O, ruthenium red, or lysine (cfompare Figures 3C and 3D to Figures 3E and 3F and Figures 4A and 4B). Direct comparisons of visualization of the glycocalyx by alcian blue or ruthenium red have suggested that alcian blue stabilizes a more extensive glycocalyx than does ruthenium red (Fassel et al 1992). Attempts to improve visualization of microbial glycocalyx by constructing cocktails of different dyes, including alcian blue and ruthenium red, alcian blue and safranin O, as well as alcian blue and lysine, were less successful than the use of individual cationic probes (data not shown).…”

Section: Discussionmentioning

confidence: 99%

High-resolution Visualization of the Microbial Glycocalyx with Low-voltage Scanning Electron Microscopy: Dependence on Cationic Dyes

Erlandsen

Kristich

Dunny

et al. 2004

J Histochem Cytochem.

147

138

View full text Add to dashboard Cite

S U M M A R YThe microbial glycocalyx is composed of a variety of polyanionic exopolysaccharides and plays important roles in microbial attachment to different substrata and to other cells. Here we report the successful use of low-voltage scanning electron microscopy (LVSEM) to visualize the glycocalyx in two microbial models ( Klebsiella pneumoniae and Enterococcus faecalis biofilms) at high resolution, and also the dependence on fixation containing polycationic dyes for its visualization. Fixation in a paraformaldehyde-glutaraldehyde cocktail without cationic dyes was inadequate for visualizing the glycocalyx, whereas addition of various dyes (alcian blue, safranin, and ruthenium red) to the aldehyde cocktail appeared necessary for stabilization. The cationic dyes varied in size, shape, and charge density, and these factors appeared responsible for different phenotypic appearances of the glycocalyx with each dye. These results suggest that aldehyde fixation with cationic dyes for high-resolution LVSEM will be a useful tool for investigation of microbial biofilms as well as investigation of the extent and role of the glycocalyx in microbial attachment to surfaces.

show abstract

Section: Discussionmentioning

confidence: 99%

High-resolution Visualization of the Microbial Glycocalyx with Low-voltage Scanning Electron Microscopy: Dependence on Cationic Dyes

Erlandsen

Kristich

Dunny

et al. 2004

J Histochem Cytochem.

147

138

View full text Add to dashboard Cite

show abstract

“…Biofilm and planktonic cells were grown on the same medium and were in stationary phase at the time of examination. The use of alcian blue is an established histochemical technique for staining and preserving polysaccharides, glycosaminoglycans, and proteoglycans in bacterial samples as well as eukaryotic cells (16). Although the quantities of uronic acids in planktonic cells and biofilms were shown to be similar, other differences in polysaccharide chemistry may account for the different distributions of the polymer around the cells.…”

Section: Discussionmentioning

confidence: 99%

Relationship between glycocalyx and povidone-iodine resistance in Pseudomonas aeruginosa (ATCC 27853) biofilms

Brown

Aldrich

Gauthier

1995

Appl Environ Microbiol

View full text Add to dashboard Cite

Biofilm-embedded bacteria are generally more resistant to antimicrobial agents than are planktonic bacteria. Two possible mechanisms for biofilm resistance are that the glycocalyx matrix secreted by cells in a biofilm reacts with and neutralizes the antimicrobial agent and that the matrix creates a diffusion barrier to the antimicrobial agent. This study was therefore conducted to examine the relationship between glycocalyx and enhanced povidone-iodine resistance in biofilms of Pseudomonas aeruginosa (ATCC 27853). Biofilms were generated by inoculation of polycarbonate membranes with broth-grown cells and incubation of them on the surfaces of nutrient agar plates. The quantities of glycocalyx material per cell were found not to be significantly different between biofilm and planktonic samples. Transmission electron microscopy showed that the distributions of glycocalyx material around cells differed in biofilm and in planktonic samples. Addition of alginic acid to planktonic cell suspensions resulted in a slight increase in resistance to povidone-iodine, suggesting some neutralizing interaction. However, the iodine demands created by biofilm and planktonic samples of equivalent biomass were not significantly different and, therefore, do not explain the contrast in resistance observed between biofilm and planktonic samples. Examination of the relationship between cell death and biomass detachment from the glycocalyx matrix revealed that most cell death occurred in the fraction of biomass that detached from a biofilm during treatment. The overall rate of iodine diffusion through biofilms was not different from that of planktonic cells collected on a polycarbonate membrane. Povidone-iodine resistance seems to be due to the protective layering of cells within the glycocalyx, which increases the time required to detach cells and, therefore, the time required for iodine to contact cells in the deepest layers of the biofilm.

show abstract

“…S 1 B). To observe the organization of the glycan layer, we fixed samples in the presence of alcian blue and lysine, which has been shown to preserve the glycan layer better (Fassel et al, 1997(Fassel et al, , 1992Fischer et al, 2012). SEM of heads of animals fixed in such a way showed that the peridermal surface was covered by a thick glycan layer, which completely obscures the underlying microridges on the head epidermis (Figure 1 B).…”

Section: Resultsmentioning

confidence: 99%

Microridges are apical projections formed of branched F-actin networks that organize the glycan layer

Khandekar

Bhavna

et al. 2018

Preprint

View full text Add to dashboard Cite

Apical projections are integral functional units of epithelial cells. Microvilli and stereocilia are cylindrical apical projections that are formed of bundled actin. Microridges on the other hand, grow laterally long, forming labyrinthine patterns on surfaces of various kinds of squamous epithelial cells. So far, the structural organization and functions of microridges have remained elusive. We have analyzed microridges on zebrafish epidermal cells using confocal and electron microscopy methods including electron tomography, to show

show abstract

Comparison of Alcian blue and ruthenium red effects on preservation of outer envelope ultrastructure in methanotrophic bacteria

Cited by 14 publications

References 36 publications

High-resolution Visualization of the Microbial Glycocalyx with Low-voltage Scanning Electron Microscopy: Dependence on Cationic Dyes

High-resolution Visualization of the Microbial Glycocalyx with Low-voltage Scanning Electron Microscopy: Dependence on Cationic Dyes

Relationship between glycocalyx and povidone-iodine resistance in Pseudomonas aeruginosa (ATCC 27853) biofilms

Microridges are apical projections formed of branched F-actin networks that organize the glycan layer

Contact Info

Product

Resources

About