13C Isotope labelLing chaMbers

Two types of labelling chamber set-ups for plants are generally used with 13C stable isotopes. The first type are commercially available, automatic cultivation chambers with a built-in regulation module for environmental parameters such as light, temperature and humidity. The second type, as oftered by Vienna Scientific, makes use of custom-made labelling devices placed in growth chambers or glasshouses which external light, temperature etc. regulation. Both approaches equally enable continuous monitoring and regulation of atmospheric parameters during labeling. Commonly conducted enrichment of other isotopes such as 15N or 34S does not require closed systems but systems allowing to add labelled liquids. Generally, both equipment types are suitable to perform pulse labeling and uniformly, long-term labeling of plants towards different isotope enrichment.

Development of 13C Isotope Labeling Chambers

The specific design of the labelling chamber will depend on the type of isotope(s) you wish to use and the specific experimental protocol you wish to follow. However, here are some general guidelines Vienna Scientific will follow to support the development of an isotope labelling chamber suiting your needs:

  • Select a chamber design: Different chamber designs are available depending on your research requirements. A typical design is a clear, airtight chamber that can hold a single plant or a group of plants. The chamber should have at least one inlet for introducing the labelled gas or solution and an outlet for removing excess gas. Other connections are beneficial.
  • Monitor and control the environment: It is important to maintain or at least monitor the temperature, humidity and CO2 concentration within the chamber. This can be achieved by using sensors and controllers to regulate the environment. Vienna Scientific can assist you in sourcing the appropriate sensors, fans, scrubbers and/or provide a design that allows existing sensors to be installed in the new chamber.
  • Build the chamber: We will then construct the chamber using PMMA or polycarbonate. The chamber will be airtight to maintain the labelled environment and will contain a positive pressure exhaust.
  • Develop the gas mixer, tubing: If more than one gas cylinder is used to supply the isotopes, an appropriate mixer must be added before the air reaches the experimental chamber. VSI will assist in the procurement of this chamber, flow meters, etc.

See the VSI webpage on ecosystem respiration chambers concerning further general recommendation for chamber designs. Contact us to discuss possible labelling chamber designs that meet your scientific objectives and budget.

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Application Potential: 13C Labelling of Rhizobox-Grown Plants

Global 13C labelling is used to produce uniformly 13C-labelled plants (here referred to as 13C plants) and requires either a continuous supply of 13CO2 throughout the cultivation or in pulses. The 13CO2 is applied as a substrate which is further converted into all the assimilates required to maintain plant metabolism. This requires airtight growth chambers (labelling chambers) with a 13CO2 atmosphere. As the production of 13C-enriched plants requires long-term cultivation, the technical equipment must allow the regulation of growth parameters such as temperature, light and humidity, as well as CO2 and O2 levels. For rhizoboxes, the rhizobox top-up chambers of Vienna Scientific allow to reach this goal cost effectvely and in combination with root and rhizosphere studies, proving a sealed top-up chamber for our "old" rhizobox design - priving an effective solution to label you rhizobox-grown plants over a longer period or in pulses. The closed chambers allow for connecting labeled gas sources and/or to connect isotopic analysers via simple in- and outlets. Advanced chamber mixing (advisable for larger top-up chambers with volumes beyond the mixing capacitiy of connected instruments) can be provided by optional internal ventilators. See the webpage on ecosystem respiration chambers concerning further recommendation for chamber designs. 

Selected Readings on 13C Labelling oF Plants

  • Abadie, C., & Tcherkez, G. (2021). 13C isotope labelling to follow the flux of photorespiratory intermediates. Plants, 10(3), 427.
  • Bromand, S., Whalen, J. K., Janzen, H. H., Schjoerring, J. K., & Ellert, B. H. (2001). A pulse-labelling method to generate 13C-enriched plant materials. Plant and Soil, 235, 253-257.
  • Ćeranić, A., Doppler, M., Büschl, C., Parich, A., Xu, K., Koutnik, A., ... & Schuhmacher, R. (2020). Preparation of uniformly labelled 13 C-and 15 N-plants using customised growth chambers. Plant Methods, 16, 1-15.
  • Hobbie, A. E., & Werner, R. A. (2004). Intramolecular, compound‐specific, and bulk carbon isotope patterns in C3 and C4 plants: a review and synthesis. New Phytologist, 161(2), 371-385.
  • Ostle, N., Ineson, P., Benham, D., & Sleep, D. (2000). Carbon assimilation and turnover in grassland vegetation using an in situ 13CO2 pulse labelling system. Rapid Communications in Mass Spectrometry, 14(15), 1345-1350.
  • Pang, R., Xu, X., Tian, Y., Cui, X., Ouyang, H., & Kuzyakov, Y. (2021). In-situ 13CO2 labeling to trace carbon fluxes in plant-soil-microorganism systems: Review and methodological guideline. Rhizosphere, 20, 100441.
  • Slaets, J. I., Resch, C., Mayr, L., Weltin, G., Heiling, M., Gruber, R., & Dercon, G. (2020). Laser spectroscopy steered 13C‐labelling of plant material in a walk‐in growth chamber. Rapid Communications in Mass Spectrometry, 34(8), e8669.
  • Soong, J. L., Reuss, D., Pinney, C., Boyack, T., Haddix, M. L., Stewart, C. E., & Cotrufo, M. F. (2014). Design and operation of a continuous 13C and 15N labeling chamber for uniform or differential, metabolic and structural, plant isotope labeling. JoVE (Journal of Visualized Experiments), (83), e51117.
  • Wang, R., Bicharanloo, B., Shirvan, M. B., Cavagnaro, T. R., Jiang, Y., Keitel, C., & Dijkstra, F. A. (2021). A novel 13C pulse‐labelling method to quantify the contribution of rhizodeposits to soil respiration in a grassland exposed to drought and nitrogen addition. New Phytologist, 230(2), 857-866.
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