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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 to multiple matrices, with increasing weights, monitored by Ultraviolet (UV) light absorption.The red regions indicate immediate recovery following incubation and the green regions indicate recovery following multiple washes. (a-d) Recovery of C60-pyrrolidine tris acid from alumina (a), VWR sand (b), wild sand (c), and natural soil (d). The results suggest that C60-pyrrolidine tris acid remains non-sticky to the various matrices.
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f5: Non-stickiness of C60-pyrrolidine tris acid to multiple matrices, with increasing weights, monitored by Ultraviolet (UV) light absorption.The red regions indicate immediate recovery following incubation and the green regions indicate recovery following multiple washes. (a-d) Recovery of C60-pyrrolidine tris acid from alumina (a), VWR sand (b), wild sand (c), and natural soil (d). The results suggest that C60-pyrrolidine tris acid remains non-sticky to the various matrices.

Mentions: The experimental setting for evaluating the non-stickiness property includes running H2O and a vacuum manifold to clear excess C60-pyrrolidine tris acid (Supplementary Figure 7). The stickiness of C60-pyrrolidine tris acid is quantified with multiple substrates, by measuring the UV absorption of filter-through at 335 nm, which is the specific absorption wavelength of C604849. With respect to synthetic substrates, alumina has been chosen as the representative substrate, since it is one of the top four components in the earth’s crust. Figure 5a indicates that more than 70% of the C60-pyrrolidine tris acid was recovered without H2O wash (red column) despite the mass of matrix. With respect to non-synthetic substrates, similar recovery rates are reported for homogenized VWR sand (>80%, Fig. 5b), Wild Sand (>60%, Fig. 5c), and Natural Soil (>70%, Fig. 5d), all without H2O wash (red column). The remainder of C60-pyrrolidine tris acid is fully recovered from the matrices, following H2O rinses (green columns in Fig. 5a–d). These results indicate that C60-pyrrolidine tris acid is not sticky to the natural environment. The rationale for requiring a second wash is due to the meso-porous architecture of the matrices that trap C60-derivatives.


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 to multiple matrices, with increasing weights, monitored by Ultraviolet (UV) light absorption.The red regions indicate immediate recovery following incubation and the green regions indicate recovery following multiple washes. (a-d) Recovery of C60-pyrrolidine tris acid from alumina (a), VWR sand (b), wild sand (c), and natural soil (d). The results suggest that C60-pyrrolidine tris acid remains non-sticky to the various matrices.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Non-stickiness of C60-pyrrolidine tris acid to multiple matrices, with increasing weights, monitored by Ultraviolet (UV) light absorption.The red regions indicate immediate recovery following incubation and the green regions indicate recovery following multiple washes. (a-d) Recovery of C60-pyrrolidine tris acid from alumina (a), VWR sand (b), wild sand (c), and natural soil (d). The results suggest that C60-pyrrolidine tris acid remains non-sticky to the various matrices.
Mentions: The experimental setting for evaluating the non-stickiness property includes running H2O and a vacuum manifold to clear excess C60-pyrrolidine tris acid (Supplementary Figure 7). The stickiness of C60-pyrrolidine tris acid is quantified with multiple substrates, by measuring the UV absorption of filter-through at 335 nm, which is the specific absorption wavelength of C604849. With respect to synthetic substrates, alumina has been chosen as the representative substrate, since it is one of the top four components in the earth’s crust. Figure 5a indicates that more than 70% of the C60-pyrrolidine tris acid was recovered without H2O wash (red column) despite the mass of matrix. With respect to non-synthetic substrates, similar recovery rates are reported for homogenized VWR sand (>80%, Fig. 5b), Wild Sand (>60%, Fig. 5c), and Natural Soil (>70%, Fig. 5d), all without H2O wash (red column). The remainder of C60-pyrrolidine tris acid is fully recovered from the matrices, following H2O rinses (green columns in Fig. 5a–d). These results indicate that C60-pyrrolidine tris acid is not sticky to the natural environment. The rationale for requiring a second wash is due to the meso-porous architecture of the matrices that trap C60-derivatives.

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.