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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.


Cellular uptake of radiotracers labelled C60-pyrrolidine tris acid by microorganisms monitored by Autoradiography.(a) and (b) 14C60-pyrrolidine tris acid uptake by E. coli (a) and B. subtilis(b). (c) and (d) 125I-C60-pyrrolidine tris acid uptake by E. coli (d) and B. subtilis (d).
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f4: Cellular uptake of radiotracers labelled C60-pyrrolidine tris acid by microorganisms monitored by Autoradiography.(a) and (b) 14C60-pyrrolidine tris acid uptake by E. coli (a) and B. subtilis(b). (c) and (d) 125I-C60-pyrrolidine tris acid uptake by E. coli (d) and B. subtilis (d).

Mentions: We then need to investigate whether C60-pyrrolidine tris acid can radiolabel microorganisms, which would thus provide a platform for imaging thick sections in an opaque environment. Even though radio tracers may have limitations in terms of resolution, having the probes for in situ imaging can motivate development of advanced instrumentation and design of novel experiments. C60-pyrrolidine tris acid is functionalized with 14C and 125I, and samples are incubated as before. Figure 4 shows β-radiation and γ-radiation emission from both E. coli (Fig. 4a,c) and B. subtilis (Fig. 4b,d), which are incubated with 14C- and 125I-labeled C60-pyrrolidine tris acid and imaged through autoradiography. The liquid scintillation data (Supplementary Figure 3) also shows a positive association of 14C-methylated C60-pyrrolidine tris acid by both E. coli and B. subtilis (about 12,000 counts per second). These results provide additional confirmation that C60-pyrrolidine tris acid can be internalized by microbes and visualized.


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 radiotracers labelled C60-pyrrolidine tris acid by microorganisms monitored by Autoradiography.(a) and (b) 14C60-pyrrolidine tris acid uptake by E. coli (a) and B. subtilis(b). (c) and (d) 125I-C60-pyrrolidine tris acid uptake by E. coli (d) and B. subtilis (d).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Cellular uptake of radiotracers labelled C60-pyrrolidine tris acid by microorganisms monitored by Autoradiography.(a) and (b) 14C60-pyrrolidine tris acid uptake by E. coli (a) and B. subtilis(b). (c) and (d) 125I-C60-pyrrolidine tris acid uptake by E. coli (d) and B. subtilis (d).
Mentions: We then need to investigate whether C60-pyrrolidine tris acid can radiolabel microorganisms, which would thus provide a platform for imaging thick sections in an opaque environment. Even though radio tracers may have limitations in terms of resolution, having the probes for in situ imaging can motivate development of advanced instrumentation and design of novel experiments. C60-pyrrolidine tris acid is functionalized with 14C and 125I, and samples are incubated as before. Figure 4 shows β-radiation and γ-radiation emission from both E. coli (Fig. 4a,c) and B. subtilis (Fig. 4b,d), which are incubated with 14C- and 125I-labeled C60-pyrrolidine tris acid and imaged through autoradiography. The liquid scintillation data (Supplementary Figure 3) also shows a positive association of 14C-methylated C60-pyrrolidine tris acid by both E. coli and B. subtilis (about 12,000 counts per second). These results provide additional confirmation that C60-pyrrolidine tris acid can be internalized by microbes and visualized.

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.