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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.


Related in: MedlinePlus

Cellular uptake of C60-pyrrolidine tris acid by E. coli and B. subtilis monitored by Transmission Electron Microscopy.The dark spots, pointed by the red arrows, represent C60-pyrrolidine-tris acid uptake by E. coli (a) and B. subtilis (b).
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f3: Cellular uptake of C60-pyrrolidine tris acid by E. coli and B. subtilis monitored by Transmission Electron Microscopy.The dark spots, pointed by the red arrows, represent C60-pyrrolidine-tris acid uptake by E. coli (a) and B. subtilis (b).

Mentions: To test the hypothesis that functionalized C60-pyrrolidine tris acid internalizes within the cell, microorganisms are imaged with TEM. Both E. coli and B. subtilis are incubated with C60-pyrrolidine tris acid, washed to remove excess compound, sectioned into slices of 60 nm thickness, and then imaged by TEM. Figure 3 shows that C60-pyrrolidine tris acid localizes within the cell in both E. coli and B. subtilis. The control experiment consists of (i) C60-pyrrolidine tris acid in DI H2O, (ii) C60-pyrrolidine tris acid on a mouse tissue section, (iii) an E. coli section without C60-pyrrolidine tris acid incubation, and (iv) a B. subtilis section without C60-pyrrolidine tris acid incubation. These data are shown in Supplementary Figure 2a–d. These results indicate positive cellular uptake of C60-derivatives in microorganisms, hence providing an opportunity to monitor the cellular activity in situ by further functionalization of C60.


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

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

Cellular uptake of C60-pyrrolidine tris acid by E. coli and B. subtilis monitored by Transmission Electron Microscopy.The dark spots, pointed by the red arrows, represent C60-pyrrolidine-tris acid uptake by E. coli (a) and B. subtilis (b).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3: Cellular uptake of C60-pyrrolidine tris acid by E. coli and B. subtilis monitored by Transmission Electron Microscopy.The dark spots, pointed by the red arrows, represent C60-pyrrolidine-tris acid uptake by E. coli (a) and B. subtilis (b).
Mentions: To test the hypothesis that functionalized C60-pyrrolidine tris acid internalizes within the cell, microorganisms are imaged with TEM. Both E. coli and B. subtilis are incubated with C60-pyrrolidine tris acid, washed to remove excess compound, sectioned into slices of 60 nm thickness, and then imaged by TEM. Figure 3 shows that C60-pyrrolidine tris acid localizes within the cell in both E. coli and B. subtilis. The control experiment consists of (i) C60-pyrrolidine tris acid in DI H2O, (ii) C60-pyrrolidine tris acid on a mouse tissue section, (iii) an E. coli section without C60-pyrrolidine tris acid incubation, and (iv) a B. subtilis section without C60-pyrrolidine tris acid incubation. These data are shown in Supplementary Figure 2a–d. These results indicate positive cellular uptake of C60-derivatives in microorganisms, hence providing an opportunity to monitor the cellular activity in situ by further functionalization of C60.

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.


Related in: MedlinePlus