Abstract
Plasmodesmata (PD) have a diameter of around 30–50 nm which is well below the 200 nm limit of optical resolution, making analysis by light microscopy difficult and resolving internal structures of the PD such as the desmotubule impossible. Modern super-resolution methods such as 3D structured illumination microscopy (3D-SIM) can increase the lateral and axial resolution and work well on fixed, sectioned material. However, imaging in live plant cells requires careful optimization. Here we present a method to image PD using 3D-SIM in live BY2 cells.
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References
Abbe E (1873) Beiträge zur Theorie des Mikroskops und der mikroskopischen Wahrnehmung. Arkiv Mikroskop Anat 9:413–468
Komis G, Šamajová O, Ovečka M, Šamaj J (2015) Super-resolution microscopy in plant cell imaging. Trends Plant Sci 20:834–843
Gustafson MGL (2000) Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy. J Microsc 198:82–87
Dobbie IM, King E, Parton RM, Carlton PM, Sedat JW, Swedlow JR, Davis I (2011) OMX: a new platform for multimodal, multi-channel wide-field imaging. Cold Spring Harb Protoc 8:899–909
Huang B, Bates M, Zhuang XW (2009) Super-resolution fluorescence microscopy. Annu Rev Biochem 78:993–1016
Fitzgibbon J, Bell K, King E, Oparka K (2010) Super-resolution imaging of plasmodesmata using 3-dimensional structured illumination microscopy. Plant Physiol 153:1453–1463
Komis G, Mistrik M, Šamajová O, Ovečka M, Bartek J, Šamaj J (2015) Superresolution live imaging of plant cells using structured illumination microscopy. Nat Protoc 10:1248–1263
Overall RL, Blackman LM (1996) A model of the macro-molecular structure of plasmodesmata. Trends Plant Sci 1:307–311
Roberts AG (2005) Plasmodesmal structure and development. In: Oparka KJ (ed) Plasmodesmata: annual plant review, vol 18. Blackwell Publishing, Oxford, pp 1–32
Nicolas WJ, Grison S, Trépout A, Gaston M, Fouché M, Cordelières FP, Oparka K, Tilsner J, Brocard L, Bayer EM (2017) Architecture and permeability of post-cytokinesis plasmodesmata lacking cytoplasmic sleeves. Nat Plants 3:17082
Tilsner J, Amari K, Torrance L (2010) Plasmodesmata viewed as specialised membrane adhesion sites. Protoplasma 248:39–36
Knox K, Wang P, Kriechbaumer V, Tilsner J, Frigerio L, Sparkes I, Hawes C, Oparka K (2015) Putting the squeeze on plasmodesmata: a role for reticulons in primary plasmodesmata formation. Plant Physiol 168:1561572
Bell K, Oparka K, Knox K (2016) Super-resolution imaging of live BY2 cells using 3D-structured illumination microscopy. Bio-protocol 6(1):e1697
Bell K, Oparka K, Knox K (2018) Preparation and imaging of specialised ER using super-resolution and TEM techniques. Methods Mol Biol 1691:33–34
Nagata T, Nemoto Y, Hasezawa S (1992) Tobacco BY-2 cell line as the "HeLa" cell in the cell biology of higher plants. Int Rev Cytol 132:1–30
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Knox, K. (2022). Super-Resolution Imaging of Plasmodesmata Using 3D Structured Illumination Microscopy. In: Benitez-Alfonso, Y., Heinlein, M. (eds) Plasmodesmata. Methods in Molecular Biology, vol 2457. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2132-5_8
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DOI: https://doi.org/10.1007/978-1-0716-2132-5_8
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