Imaging the Root Hair Morphology of Arabidopsis Seedlings in a Two-layer Microfluidic Platform
Abstract
Root hairs increase root surface area for better water uptake and nutrient absorption by the plant. Because they are small in size and often obscured by their natural environment, root hair morphology and function are difficult to study and often excluded from plant research. In recent years, microfluidic platforms have offered a way to visualize root systems at high resolution without disturbing the roots during transfer to an imaging system. The microfluidic platform presented here builds on previous plant-on-a-chip research by incorporating a two-layer device to confine the Arabidopsis thaliana main root to the same optical plane as the root hairs. This design enables the quantification of root hairs on a cellular and organelle level and also prevents z-axis drifting during the addition of experimental treatments. We describe how to store the devices in a contained and hydrated environment, without the need for fluidic pumps, while maintaining a gnotobiotic environment for the seedling. After the optical imaging experiment, the device may be disassembled and used as a substrate for atomic force or scanning electron microscopy while keeping fine root structures intact.
- Authors:
-
- Univ. of Tennessee, Knoxville, TN (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1427673
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Visualized Experiments
- Additional Journal Information:
- Journal Volume: 126; Journal Issue: 126; Journal ID: ISSN 1940-087X
- Publisher:
- MyJoVE Corp.
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Aufrecht, Jayde A., Ryan, Jennifer M., Hasim, Sahar, Allison, David P., Nebenführ, Andreas, Doktycz, Mitchel J., and Retterer, Scott T. Imaging the Root Hair Morphology of Arabidopsis Seedlings in a Two-layer Microfluidic Platform. United States: N. p., 2017.
Web. doi:10.3791/55971.
Aufrecht, Jayde A., Ryan, Jennifer M., Hasim, Sahar, Allison, David P., Nebenführ, Andreas, Doktycz, Mitchel J., & Retterer, Scott T. Imaging the Root Hair Morphology of Arabidopsis Seedlings in a Two-layer Microfluidic Platform. United States. https://doi.org/10.3791/55971
Aufrecht, Jayde A., Ryan, Jennifer M., Hasim, Sahar, Allison, David P., Nebenführ, Andreas, Doktycz, Mitchel J., and Retterer, Scott T. Tue .
"Imaging the Root Hair Morphology of Arabidopsis Seedlings in a Two-layer Microfluidic Platform". United States. https://doi.org/10.3791/55971. https://www.osti.gov/servlets/purl/1427673.
@article{osti_1427673,
title = {Imaging the Root Hair Morphology of Arabidopsis Seedlings in a Two-layer Microfluidic Platform},
author = {Aufrecht, Jayde A. and Ryan, Jennifer M. and Hasim, Sahar and Allison, David P. and Nebenführ, Andreas and Doktycz, Mitchel J. and Retterer, Scott T.},
abstractNote = {Root hairs increase root surface area for better water uptake and nutrient absorption by the plant. Because they are small in size and often obscured by their natural environment, root hair morphology and function are difficult to study and often excluded from plant research. In recent years, microfluidic platforms have offered a way to visualize root systems at high resolution without disturbing the roots during transfer to an imaging system. The microfluidic platform presented here builds on previous plant-on-a-chip research by incorporating a two-layer device to confine the Arabidopsis thaliana main root to the same optical plane as the root hairs. This design enables the quantification of root hairs on a cellular and organelle level and also prevents z-axis drifting during the addition of experimental treatments. We describe how to store the devices in a contained and hydrated environment, without the need for fluidic pumps, while maintaining a gnotobiotic environment for the seedling. After the optical imaging experiment, the device may be disassembled and used as a substrate for atomic force or scanning electron microscopy while keeping fine root structures intact.},
doi = {10.3791/55971},
journal = {Journal of Visualized Experiments},
number = 126,
volume = 126,
place = {United States},
year = {Tue Aug 01 00:00:00 EDT 2017},
month = {Tue Aug 01 00:00:00 EDT 2017}
}
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Works referencing / citing this record:
Label-free time- and space-resolved exometabolite sampling of growing plant roots through nanoporous interfaces
journal, July 2019
- Patabadige, Damith E. W.; Millet, Larry J.; Aufrecht, Jayde A.
- Scientific Reports, Vol. 9, Issue 1
Increasing access to microfluidics for studying fungi and other branched biological structures
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- Millet, Larry J.; Aufrecht, Jayde; Labbé, Jessy
- Fungal Biology and Biotechnology, Vol. 6, Issue 1