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Functionalized Buckyballs for Visualizing Microbial Species in Different States and Environments.

Cheng Q, Aravind A, Buckley M, Gifford A, Parvin B - Sci Rep (2015)

Bottom Line: To date, in situ visualization of microbial density has remained an open problem.Here, functionalized buckyballs (e.g., C60-pyrrolidine tris acid) are shown to be a versatile platform that allows internalization within a microorganism without either adhering to the cell wall and cell membrane or binding to a matrix substrate such as soil.We also demonstrate that cysteine-functionalized C60-pyrrolidine tris acid can differentiate live and dead microorganisms.

View Article: PubMed Central - PubMed

Affiliation: Department of Electrical and Biomedical Engineering, University of Nevada, Reno, 1664 N Virginia Street, Reno NV, 89503, USA.

ABSTRACT
To date, in situ visualization of microbial density has remained an open problem. Here, functionalized buckyballs (e.g., C60-pyrrolidine tris acid) are shown to be a versatile platform that allows internalization within a microorganism without either adhering to the cell wall and cell membrane or binding to a matrix substrate such as soil. These molecular probes are validated via multi-scale imaging, to show association with microorganisms via fluorescence microscopy, positive cellular uptake via electron microscopy, and non-specific binding to the substrates through a combination of fluorescence and autoradiography imaging. We also demonstrate that cysteine-functionalized C60-pyrrolidine tris acid can differentiate live and dead microorganisms.

No MeSH data available.


Non-stickiness of C60-pyrrolidine tris acid monitored by Confocal Laser Scanning Microscopy following several washes.Fluorescence microscopy indicates that fBSA labelled C60-pyrrolidine tris acid has no adherence to alumina (a) and glass beads (b), which are clearly present in bright field microscopy.
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f6: Non-stickiness of C60-pyrrolidine tris acid monitored by Confocal Laser Scanning Microscopy following several washes.Fluorescence microscopy indicates that fBSA labelled C60-pyrrolidine tris acid has no adherence to alumina (a) and glass beads (b), which are clearly present in bright field microscopy.

Mentions: To investigate the non-stickiness property of C60-pyrrolidine tris acid spatially, several experiments have been designed. (i) fBSA-labeled C60-pyrrolidine tris acid is evaluated against non-auto-fluorescent matrices such as glass beads and alumina (Fig. 6). Using fluorescence microscopy, each substrate emits an initial fluorescent signal after incubation with C60-pyrrolidine tris-fBSA, with the signal being lost following multiple H2O washes, thus providing confirmation that C60-pyrrolidine was not sticky to the natural environment. (ii) fBSA-labeled C60-pyrrolidine tris acid is evaluated against matrices such as VWR sand, wild sand, and natural soil (Supplementary Figure 8). However, these matrices are auto-fluorescent and mask fluorescent probes, making it difficult to visualize the fluorescent-labeled C60. (iii) Further validation by autoradiography and liquid scintillation is pursued in all substrates (e.g., pretreated VWR sand, glass beads, alumina, wild sand, and natural soil). Figure 7 indicates that neither β- nor γ-radiation are detected from incubated matrices (middle and right columns) after 6 H2O rinses. Therefore, we conclude that C60-derivatives are non-sticky to the natural environment and can be removed entirely by H2O washes.


Functionalized Buckyballs for Visualizing Microbial Species in Different States and Environments.

Cheng Q, Aravind A, Buckley M, Gifford A, Parvin B - Sci Rep (2015)

Non-stickiness of C60-pyrrolidine tris acid monitored by Confocal Laser Scanning Microscopy following several washes.Fluorescence microscopy indicates that fBSA labelled C60-pyrrolidine tris acid has no adherence to alumina (a) and glass beads (b), which are clearly present in bright field microscopy.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561912&req=5

f6: Non-stickiness of C60-pyrrolidine tris acid monitored by Confocal Laser Scanning Microscopy following several washes.Fluorescence microscopy indicates that fBSA labelled C60-pyrrolidine tris acid has no adherence to alumina (a) and glass beads (b), which are clearly present in bright field microscopy.
Mentions: To investigate the non-stickiness property of C60-pyrrolidine tris acid spatially, several experiments have been designed. (i) fBSA-labeled C60-pyrrolidine tris acid is evaluated against non-auto-fluorescent matrices such as glass beads and alumina (Fig. 6). Using fluorescence microscopy, each substrate emits an initial fluorescent signal after incubation with C60-pyrrolidine tris-fBSA, with the signal being lost following multiple H2O washes, thus providing confirmation that C60-pyrrolidine was not sticky to the natural environment. (ii) fBSA-labeled C60-pyrrolidine tris acid is evaluated against matrices such as VWR sand, wild sand, and natural soil (Supplementary Figure 8). However, these matrices are auto-fluorescent and mask fluorescent probes, making it difficult to visualize the fluorescent-labeled C60. (iii) Further validation by autoradiography and liquid scintillation is pursued in all substrates (e.g., pretreated VWR sand, glass beads, alumina, wild sand, and natural soil). Figure 7 indicates that neither β- nor γ-radiation are detected from incubated matrices (middle and right columns) after 6 H2O rinses. Therefore, we conclude that C60-derivatives are non-sticky to the natural environment and can be removed entirely by H2O washes.

Bottom Line: To date, in situ visualization of microbial density has remained an open problem.Here, functionalized buckyballs (e.g., C60-pyrrolidine tris acid) are shown to be a versatile platform that allows internalization within a microorganism without either adhering to the cell wall and cell membrane or binding to a matrix substrate such as soil.We also demonstrate that cysteine-functionalized C60-pyrrolidine tris acid can differentiate live and dead microorganisms.

View Article: PubMed Central - PubMed

Affiliation: Department of Electrical and Biomedical Engineering, University of Nevada, Reno, 1664 N Virginia Street, Reno NV, 89503, USA.

ABSTRACT
To date, in situ visualization of microbial density has remained an open problem. Here, functionalized buckyballs (e.g., C60-pyrrolidine tris acid) are shown to be a versatile platform that allows internalization within a microorganism without either adhering to the cell wall and cell membrane or binding to a matrix substrate such as soil. These molecular probes are validated via multi-scale imaging, to show association with microorganisms via fluorescence microscopy, positive cellular uptake via electron microscopy, and non-specific binding to the substrates through a combination of fluorescence and autoradiography imaging. We also demonstrate that cysteine-functionalized C60-pyrrolidine tris acid can differentiate live and dead microorganisms.

No MeSH data available.