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Title: Label-free time- and space-resolved exometabolite sampling of growing plant roots through nanoporous interfaces

Abstract

Spatial and temporal profiling of metabolites within and between living systems is vital to understanding how chemical signaling shapes the composition and function of these complex systems. Measurement of metabolites is challenging because they are often not amenable to extrinsic tags, are diverse in nature, and are present with a broad range of concentrations. Moreover, direct imaging by chemically informative tools can significantly compromise viability of the system of interest or lack adequate resolution. The development of technologies to “image” metabolites is essential to provide spatiotemporal resolution that cannot be obtained by previous approaches. Here, we present a nano-enabled and label-free imaging technology using a microfluidic sampling network to track production and distribution of chemical information in the microenvironment of a living organism. We demonstrate the sampling of exometabolites secreted from a growing plant over time, providing spatial and temporal information about development within a model environment. We describe two approaches that integrate a polyester track-etched (PETE) nanofluidic interface to physically confine the biological sample within the model environment, while allowing fluidic access via an underlying microfluidic network. The nanoporous interface enables sampling of the microenvironment above in a time-dependent and spatially-resolved manner. Chemical information was extracted at two differentmore » locations, relative to the growing plant root, over time. The flow of metabolites was controlled by tuning the sample aperture size or “pixel size” of the patterned PETE membrane. The diffusional flux through the PETE membrane was characterized to understand membrane performance. Exometabolites of a growing plant root were successfully profiled in a space- and time-resolved manner. This method and device provide a frame-by-frame description of the chemical environment that maps to the physical and biological characteristics of the sample.« less

Authors:
; ; ; ; ; ; ORCiD logo
Publication Date:
Research Org.:
Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1619569
Alternate Identifier(s):
OSTI ID: 1543220
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Published Article
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Name: Scientific Reports Journal Volume: 9 Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United Kingdom
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Patabadige, Damith E. W., Millet, Larry J., Aufrecht, Jayde A., Shankles, Peter G., Standaert, Robert F., Retterer, Scott T., and Doktycz, Mitchel J. Label-free time- and space-resolved exometabolite sampling of growing plant roots through nanoporous interfaces. United Kingdom: N. p., 2019. Web. doi:10.1038/s41598-019-46538-5.
Patabadige, Damith E. W., Millet, Larry J., Aufrecht, Jayde A., Shankles, Peter G., Standaert, Robert F., Retterer, Scott T., & Doktycz, Mitchel J. Label-free time- and space-resolved exometabolite sampling of growing plant roots through nanoporous interfaces. United Kingdom. https://doi.org/10.1038/s41598-019-46538-5
Patabadige, Damith E. W., Millet, Larry J., Aufrecht, Jayde A., Shankles, Peter G., Standaert, Robert F., Retterer, Scott T., and Doktycz, Mitchel J. Tue . "Label-free time- and space-resolved exometabolite sampling of growing plant roots through nanoporous interfaces". United Kingdom. https://doi.org/10.1038/s41598-019-46538-5.
@article{osti_1619569,
title = {Label-free time- and space-resolved exometabolite sampling of growing plant roots through nanoporous interfaces},
author = {Patabadige, Damith E. W. and Millet, Larry J. and Aufrecht, Jayde A. and Shankles, Peter G. and Standaert, Robert F. and Retterer, Scott T. and Doktycz, Mitchel J.},
abstractNote = {Spatial and temporal profiling of metabolites within and between living systems is vital to understanding how chemical signaling shapes the composition and function of these complex systems. Measurement of metabolites is challenging because they are often not amenable to extrinsic tags, are diverse in nature, and are present with a broad range of concentrations. Moreover, direct imaging by chemically informative tools can significantly compromise viability of the system of interest or lack adequate resolution. The development of technologies to “image” metabolites is essential to provide spatiotemporal resolution that cannot be obtained by previous approaches. Here, we present a nano-enabled and label-free imaging technology using a microfluidic sampling network to track production and distribution of chemical information in the microenvironment of a living organism. We demonstrate the sampling of exometabolites secreted from a growing plant over time, providing spatial and temporal information about development within a model environment. We describe two approaches that integrate a polyester track-etched (PETE) nanofluidic interface to physically confine the biological sample within the model environment, while allowing fluidic access via an underlying microfluidic network. The nanoporous interface enables sampling of the microenvironment above in a time-dependent and spatially-resolved manner. Chemical information was extracted at two different locations, relative to the growing plant root, over time. The flow of metabolites was controlled by tuning the sample aperture size or “pixel size” of the patterned PETE membrane. The diffusional flux through the PETE membrane was characterized to understand membrane performance. Exometabolites of a growing plant root were successfully profiled in a space- and time-resolved manner. This method and device provide a frame-by-frame description of the chemical environment that maps to the physical and biological characteristics of the sample.},
doi = {10.1038/s41598-019-46538-5},
journal = {Scientific Reports},
number = 1,
volume = 9,
place = {United Kingdom},
year = {Tue Jul 16 00:00:00 EDT 2019},
month = {Tue Jul 16 00:00:00 EDT 2019}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1038/s41598-019-46538-5

Citation Metrics:
Cited by: 8 works
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Figures / Tables:

Figure 1 Figure 1: Microfluidic device for simultaneous imaging of live root development and for metabolite sampling. (A) Schematic showing assembly of the three layers used to produce a culture system for a growing wheat root, infusion-patterned nanoporous membrane for sampling, and sample collection for analysis of metabolites. (B) Photograph of amore » germinated wheat seed growing in the device. (C) The primary root is placed in the sample microenvironment layer. The underlying sampling channels (C1 and C2) allow fluid pumping through ports, enabling buffer exchange and sampling of metabolites at two individually accessible locations. (D) Brightfield image showing the plant root in the primary microchannel after 6 h of growth in the device. The underlying metabolite sampling channels, highlighted by dashed lines, are obscured by the sandwiched nanoporous membrane. (E) Mass spectra derived from extracted-ion chromatogram (XIC) of sucrose indicating differential levels of sucrose in samples collected proximal and distal to the seed. The trimethylsilyl derivative of sucrose elutes at 15.09 min under the given separation conditions. The sucrose chromatographic peaks were determined by aligning with the given XIC for a sucrose standard. The Student t-test indicates that sucrose produced at the C1 and C2 locations are significantly different at P < 0.05 (95% confidence interval).« less

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