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Chloroplast Biotechnology pp 125–143Cite as

Fluorescent Labeling and Confocal Microscopic Imaging of Chloroplasts and Non-green Plastids

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1132))

Abstract

While chlorophyll has served as an excellent label for plastids in green tissue, the development of fluorescent proteins has allowed their ready visualization in all tissues of the plants, revealing new features of their morphology and motility. Gene regulatory sequences in plastid transgenes can be optimized through the use of fluorescent protein reporters. Fluorescent labeling of plastids simultaneously with other subcellular locations reveals dynamic interactions and mutant phenotypes. Transient expression of fluorescent protein fusions is particularly valuable to determine whether or not a protein of unknown function is targeted to the plastid. Particle bombardment and agroinfiltration methods described here are convenient for imaging fluorescent proteins in plant organelles. With proper selection of fluorophores for labeling the components of the plant cell, confocal microscopy can produce extremely informative images at high resolution at depths not feasible by standard epifluorescence microscopy.

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Acknowledgments

This work was supported by grants from the Chemical Sciences, Geosciences, and Biosciences Division, Office of Basic Energy Sciences, Office of Science, US Department of Energy to M.R.H., including DE-89-ER14030 and DE–FG02–09ER16070.

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Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA

    Maureen R. Hanson & Amirali Sattarzadeh

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  1. Maureen R. Hanson
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  1. Rutgers University, Waksman Inst.of Microbiology, Piscataway, New Jersey, USA

    Pal Maliga

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Hanson, M.R., Sattarzadeh, A. (2014). Fluorescent Labeling and Confocal Microscopic Imaging of Chloroplasts and Non-green Plastids. In: Maliga, P. (eds) Chloroplast Biotechnology. Methods in Molecular Biology, vol 1132. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-995-6_7

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