Abstract
The high-throughput analysis of respiratory activity has become an important component of many biological investigations. Here, a technological platform, denoted the “MultiSense tool,” is described. The tool enables the parallel monitoring of respiration in 100 samples over an extended time period, by dynamically tracking the concentrations of oxygen (O2) and/or carbon dioxide (CO2) and/or pH within an airtight vial. Its flexible design supports the quantification of respiration based on either oxygen consumption or carbon dioxide release, thereby allowing for the determination of the physiologically significant respiratory quotient (the ratio between the quantities of CO2 released and the O2 consumed). It requires an LED light source to be mounted above the sample, together with a CCD camera system, adjusted to enable the capture of analyte-specific wavelengths, and fluorescent sensor spots inserted into the sample vial. Here, a demonstration is given of the use of the MultiSense tool to quantify respiration in imbibing plant seeds, for which an appropriate step-by-step protocol is provided. The technology can be easily adapted for a wide range of applications, including the monitoring of gas exchange in any kind of liquid culture system (algae, embryo and tissue culture, cell suspensions, microbial cultures).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Van Asbrouck J, Taridno P (2009) Using the single seed oxygen consumption measurement as a method of determination of different seed quality parameters for commercial tomato seed samples. Asian J Food Agro-Ind 2:S88–S95
Sew YS, Ströher E, Holzmann C et al (2013) Multiplex micro-respiratory measurements of Arabidopsis tissues. New Phytol 200:922–932
Xin X, Wan Y, Wang W et al (2013) A real-time, non-invasive, micro-optrode technique for detecting seed viability by using oxygen influx. Sci Rep 3:3057
Millar A.H, Siedow J.N., Day D. (2015) Respiration and photorespiration. In: B.B. Buchanan, W. Gruissem, and R.L. Jones. Biochemistry and molecular biology of plants, 2, John Wiley & Sons Ltd, New York pp. 610–655.
Meier R, Schreml S, Wang X-D et al (2011) Simultaneous photographing of oxygen and pH in vivo using sensor films. Angew Chem Int Ed 50:10893–10896
Wang X-D, Meier RJ, Link M et al (2010) Photographing oxygen distribution. Angew Chem Int Ed 49:4907–4909
Wang X-D, Meier RJ, Wolfbeis OS (2013) Fluorescent pH-sensitive nanoparticles in an Agarose matrix for imaging of bacterial growth and metabolism. Angew Chem Int Ed 52:406–409
Blossfeld S, Schreiber CM, Liebsch G et al (2013) Quantitative imaging of rhizosphere pH and CO2 dynamics with planar optodes. Ann Bot 112:267–276
Rolletschek H., Liebsch G. (this issue). A method for imaging oxygen distribution and respiration at a microscopic level of resolution. MiMB
Tschiersch H, Liebsch G, Borisjuk L et al (2014) Imaging microbial culture O2 consumption. Genet Eng Biotechnol News 34:30
Stoll VS, Blanchard JS (2009) Buffers: principles and practice. Methods Enzymol 463:43–56
Schwender J, Hebbelmann I, Heinzel N et al (2015) Quantitative multilevel analysis of central metabolism in developing oilseeds of Brassica napus during in vitro culture. Plant Physiol 168:828–848
Borisjuk L, Neuberger T, Schwender J et al (2013) Seed architecture shapes embryo metabolism in oilseed rape. Plant Cell 25:1625–1640
Sauer DB, Burroughs R (1986) Disinfection of seed surfaces with sodium hypochlorite. Phytopathology 76:745–749
Lin RZ, Chang HY (2008) Recent advances in three-dimensional multicellular spheroid culture for biomedical research. Biotechnol J 3(9–10):1172–1184
S W, Doetsch J, Mueller-Klieser W, Kunz-Schughart LA (2000) Metabolic imaging in multicellular spheroids of oncogene-transfected fibroblasts. J Histochem Cytochem 48(4):509–522
Gromosova EN, Fomina MA, Blazchuk JS (1995) Development peculiarities of mycelial structures of Thielavia terrestris under batch conditions. Biopolym Cell 11(3–4):73–81
Chang ST, Miles PG (2004) Cultivation, nutritional value, medicinal effect and environmental impact of mushrooms. CRC Press, Boca Raton, pp 73–77. ISBN 9780849310430
Shen X, Gmitter FG Jr, Grosser JW (2011) Immature embryo rescue and culture. Methods Mol Biol 710:75–92
Acknowledgements
This research was financially supported by the German Plant Phenotyping Network (DPPN).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Keil, P., Liebsch, G., Borisjuk, L., Rolletschek, H. (2017). MultiSense: A Multimodal Sensor Tool Enabling the High-Throughput Analysis of Respiration. In: Jagadis Gupta, K. (eds) Plant Respiration and Internal Oxygen. Methods in Molecular Biology, vol 1670. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7292-0_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7292-0_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7291-3
Online ISBN: 978-1-4939-7292-0
eBook Packages: Springer Protocols