Critical steps and troubleshooting in sample preparation for wood and phloem formation: from sampling to microscopic observation
Kritični koraki in reševanje težav pri pripravi vzorcev za spremljanje nastajanja lesa in floema: od vzorčenja do opazovanja pod mikroskopom
DOI:
https://doi.org/10.26614/les-wood.2022.v71n01a05Keywords:
tissue sampling, tissue preparation, microscopic slides, microscopy, xylogenesis, phloem formation, troubleshootingAbstract
We present a technical note that supplements published procedures on optimal sample preparation for performing wood and phloem formation analyses. Before beginning sampling, it is important to learn about the characteristics of the tree or shrub species to be investigated. Some tips are given how to use the Trephor tool in the best way, how to remove the outer hard bark (periderm), how microcores should be handled after removal from the tree, and how they should be oriented for embedding in paraffin, and cutting thin sections for microscopy. Possible defects that may result from improper handling are illustrated and discussed. We also present optimal images to accurately identify different cell development stages in phloem and xylem, which is particularly challenging in hardwoods and Mediterranean tree and shrub species.
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Balzano, A., Čufar, K., & De Micco, V. (2021a). Cell-wall fluorescence highlights the phases of xylogenesis. IAWA Journal, 43 (1, 2), 80-91, DOI: https://doi.org/10.1163/22941932-bja10080
Balzano, A., Čufar, K., & De Micco, V. (2021b). Xylem and phloem formation dynamics in Quercus ilex L. at a dry site in Southern Italy. Forests, 12(2), 188, DOI: https://doi.org/10.3390/f12020188
De Micco, V., Carrer, M., Rathgeber, C. B., Camarero, J. J., Voltas, J., Cherubini, P., & Battipaglia, G. (2019). From xylogenesis to tree rings: wood traits to investigate tree response to environmental changes. IAWA journal, 40(2), 155-182, DOI: https://doi.org/10.1163/22941932-40190246
Deslauriers, A., Rossi, S., & Liang, E. (2015). Collecting and processing wood microcores for monitoring xylogenesis. In Plant microtechniques and protocols (pp. 417-429). Springer, Cham. DOI: https://doi.org/10.1007/978-3-319-19944-3_23
Pace, M. R. (2019). Optimal preparation of tissue sections for light-microscopic analysis of phloem anatomy. In Phloem (pp. 3-16). Humana, New York, NY. DOI: https://doi.org/10.1007/978-1-4939-9562-2_1
Prislan, P., Gričar, J., & Čufar, K. (2014a). Wood sample preparation for microscopic analysis – protocol. URL: http://www.streess-cost.eu/images/stories/Documents/protocol_wood_sample_preparation_for_microscopic_analysis.pdf
Prislan, P., Martinez del Castillo, E., Krže, L., Habjan, P., & Merela, M. (2014b). Wood sample preparation for microscopic analysis film. URL: https://www.youtube.com/watch?v=FUddCGVr0bY
Prislan, P., Martinez del Castillo, E., Skoberne, G., Špenko, N., & Gričar, J. (2022). Sample preparation protocol for wood and phloem formation analyses. Dendrochronologia, 73, 125959, DOI: https://doi.org/10.1016/j.dendro.2022.125959
Rossi, S., Anfodillo, T., & Menardi, R. (2006). Trephor: a new tool for sampling microcores from tree stems. Iawa Journal, 27(1), 89-97, DOI: https://doi.org/10.1163/22941932-90000139
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