Time-lapse Fluorescence Imaging of Arabidopsis Root Growth with Rapid Manipulation of The Root Environment Using The RootChip
Abstract
The root functions as the physical anchor of the plant and is the organ responsible for uptake of water and mineral nutrients such as nitrogen, phosphorus, sulfate and trace elements that plants acquire from the soil. If we want to develop sustainable approaches to producing high crop yield, we need to better understand how the root develops, takes up a wide spectrum of nutrients, and interacts with symbiotic and pathogenic organisms. To accomplish these goals, we need to be able to explore roots in microscopic detail over time periods ranging from minutes to days. We developed the RootChip, a polydimethylsiloxane (PDMS)- based microfluidic device, which allows us to grow and image roots from Arabidopsis seedlings while avoiding any physical stress to roots during preparation for imaging1 (Figure 1). The device contains a bifurcated channel structure featuring micromechanical valves to guide the fluid flow from solution inlets to each of the eight observation chambers2 . This perfusion system allows the root microenvironment to be controlled and modified with precision and speed. The volume of the chambers is approximately 400 nl, thus requiring only minimal amounts of test solution. Here we provide a detailed protocol for studying root biology on the RootChipmore »
- Authors:
-
- Stanford Univ., CA (United States). Carnegie Inst. for Science. Dept. of Plant Biology
- Howard Hughes Medical Inst., Chevy Chase, MD (United States); Stanford Univ., CA (United States). Dept. of Appleid Physics and Bioengineering; Univ. of Freiburg (Germany). Center for Biological Signaling Studies (BIOSS). Dept. of Microsystems Engineering (IMTEK)
- Howard Hughes Medical Inst., Chevy Chase, MD (United States); Stanford Univ., CA (United States). Dept. of Appleid Physics and Bioengineering
- Publication Date:
- Research Org.:
- Carnegie Inst. of Washington, Washington, DC (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division; National Science Foundation (NSF)
- OSTI Identifier:
- 1628632
- Grant/Contract Number:
- FG02-04ER15542; MCB 1021677
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Visualized Experiments
- Additional Journal Information:
- Journal Issue: 65; Journal ID: ISSN 1940-087X
- Publisher:
- MyJoVE Corp.
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 77 NANOSCIENCE AND NANOTECHNOLOGY; science & technology; bioengineering; plant biology; physics; plant physiology; roots; microfluidics; imaging; hydroponics; Arabidopsis
Citation Formats
Grossmann, Guido, Meier, Matthias, Cartwright, Heather N., Sosso, Davide, Quake, Stephen R., Ehrhardt, David W., and Frommer, Wolf B. Time-lapse Fluorescence Imaging of Arabidopsis Root Growth with Rapid Manipulation of The Root Environment Using The RootChip. United States: N. p., 2012.
Web. doi:10.3791/4290.
Grossmann, Guido, Meier, Matthias, Cartwright, Heather N., Sosso, Davide, Quake, Stephen R., Ehrhardt, David W., & Frommer, Wolf B. Time-lapse Fluorescence Imaging of Arabidopsis Root Growth with Rapid Manipulation of The Root Environment Using The RootChip. United States. https://doi.org/10.3791/4290
Grossmann, Guido, Meier, Matthias, Cartwright, Heather N., Sosso, Davide, Quake, Stephen R., Ehrhardt, David W., and Frommer, Wolf B. Sat .
"Time-lapse Fluorescence Imaging of Arabidopsis Root Growth with Rapid Manipulation of The Root Environment Using The RootChip". United States. https://doi.org/10.3791/4290. https://www.osti.gov/servlets/purl/1628632.
@article{osti_1628632,
title = {Time-lapse Fluorescence Imaging of Arabidopsis Root Growth with Rapid Manipulation of The Root Environment Using The RootChip},
author = {Grossmann, Guido and Meier, Matthias and Cartwright, Heather N. and Sosso, Davide and Quake, Stephen R. and Ehrhardt, David W. and Frommer, Wolf B.},
abstractNote = {The root functions as the physical anchor of the plant and is the organ responsible for uptake of water and mineral nutrients such as nitrogen, phosphorus, sulfate and trace elements that plants acquire from the soil. If we want to develop sustainable approaches to producing high crop yield, we need to better understand how the root develops, takes up a wide spectrum of nutrients, and interacts with symbiotic and pathogenic organisms. To accomplish these goals, we need to be able to explore roots in microscopic detail over time periods ranging from minutes to days. We developed the RootChip, a polydimethylsiloxane (PDMS)- based microfluidic device, which allows us to grow and image roots from Arabidopsis seedlings while avoiding any physical stress to roots during preparation for imaging1 (Figure 1). The device contains a bifurcated channel structure featuring micromechanical valves to guide the fluid flow from solution inlets to each of the eight observation chambers2 . This perfusion system allows the root microenvironment to be controlled and modified with precision and speed. The volume of the chambers is approximately 400 nl, thus requiring only minimal amounts of test solution. Here we provide a detailed protocol for studying root biology on the RootChip using imaging-based approaches with real time resolution. Roots can be analyzed over several days using time lapse microscopy. Roots can be perfused with nutrient solutions or inhibitors, and up to eight seedlings can be analyzed in parallel. This system has the potential for a wide range of applications, including analysis of root growth in the presence or absence of chemicals, fluorescence-based analysis of gene expression, and the analysis of biosensors, e.g. FRET nanosensors3.},
doi = {10.3791/4290},
journal = {Journal of Visualized Experiments},
number = 65,
volume = ,
place = {United States},
year = {Sat Jul 07 00:00:00 EDT 2012},
month = {Sat Jul 07 00:00:00 EDT 2012}
}
Works referenced in this record:
GLUT1 and GLUT9 as major contributors to glucose influx in HepG2 cells identified by a high sensitivity intramolecular FRET glucose sensor
journal, April 2008
- Takanaga, Hitomi; Chaudhuri, Bhavna; Frommer, Wolf B.
- Biochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1778, Issue 4
Imaging approach for monitoring cellular metabolites and ions using genetically encoded biosensors
journal, February 2010
- Okumoto, Sakiko
- Current Opinion in Biotechnology, Vol. 21, Issue 1
Solvent Compatibility of Poly(dimethylsiloxane)-Based Microfluidic Devices
journal, October 2003
- Lee, Jessamine Ng; Park, Cheolmin; Whitesides, George M.
- Analytical Chemistry, Vol. 75, Issue 23, p. 6544-6554
The origins and the future of microfluidics
journal, July 2006
- Whitesides, George M.
- Nature, Vol. 442, Issue 7101, p. 368-373
A cytosolic trans-activation domain essential for ammonium uptake
journal, February 2007
- Loqué, D.; Lalonde, S.; Looger, L. L.
- Nature, Vol. 446, Issue 7132
Chemical stimulation of the Arabidopsis thaliana root using multi-laminar flow on a microfluidic chip
journal, January 2010
- Meier, Matthias; Lucchetta, Elena M.; Ismagilov, Rustem F.
- Lab on a Chip, Vol. 10, Issue 16
Imaging of metabolites by using a fusion protein between a periplasmic binding protein and GFP derivatives: From a chimera to a view of reality
journal, July 2002
- Stitt, Mark
- Proceedings of the National Academy of Sciences, Vol. 99, Issue 15
Dynamic imaging of glucose flux impedance using FRET sensors in wild-type Arabidopsis plants
journal, January 2011
- Chaudhuri, B.; Hormann, F.; Frommer, W. B.
- Journal of Experimental Botany, Vol. 62, Issue 7
The RootChip: An Integrated Microfluidic Chip for Plant Science
journal, December 2011
- Grossmann, Guido; Guo, Woei-Jiun; Ehrhardt, David W.
- The Plant Cell, Vol. 23, Issue 12
Monolithic Microfabricated Valves and Pumps by Multilayer Soft Lithography
journal, April 2000
- Unger, M. A.
- Science, Vol. 288, Issue 5463
Plant-in-chip: Microfluidic system for studying root growth and pathogenic interactions in Arabidopsis
dataset, January 2022
- Pandey, Santosh
- Harvard Dataverse
Works referencing / citing this record:
Multiscale and Multimodal Approaches to Study Autophagy in Model Plants
journal, January 2018
- Marion, Jessica; Le Bars, Romain; Besse, Laetitia
- Cells, Vol. 7, Issue 1
Soil-on-a-Chip: microfluidic platforms for environmental organismal studies
journal, January 2016
- Stanley, Claire E.; Grossmann, Guido; Casadevall i. Solvas, Xavier
- Lab on a Chip, Vol. 16, Issue 2
Multiple cyclic nucleotide‐gated channels coordinate calcium oscillations and polar growth of root hairs
journal, June 2019
- Brost, Christa; Studtrucker, Tanja; Reimann, Ronny
- The Plant Journal
Male–female communication triggers calcium signatures during fertilization in Arabidopsis
journal, August 2014
- Denninger, Philipp; Bleckmann, Andrea; Lausser, Andreas
- Nature Communications, Vol. 5, Issue 1
Rapid and reversible root growth inhibition by TIR1 auxin signalling
journal, June 2018
- Fendrych, Matyáš; Akhmanova, Maria; Merrin, Jack
- Nature Plants, Vol. 4, Issue 7
Multiscale and Multimodal Approaches to Study Autophagy in Model Plants
journal, January 2018
- Marion, Jessica; Le Bars, Romain; Besse, Laetitia
- Cells, Vol. 7, Issue 1
What Has Been Seen Cannot Be Unseen—Detecting Auxin In Vivo
journal, December 2017
- Pařízková, Barbora; Pernisová, Markéta; Novák, Ondřej
- International Journal of Molecular Sciences, Vol. 18, Issue 12