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

Authors

  • Angela Balzano
  • Katarina Čufar
  • Luka Krže
  • Maks Merela

DOI:

https://doi.org/10.26614/les-wood.2022.v71n01a05

Keywords:

tissue sampling, tissue preparation, microscopic slides, microscopy, xylogenesis, phloem formation, troubleshooting

Abstract

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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References

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 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 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 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 DOI: https://doi.org/10.1007/978-3-319-19944-3_23

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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 DOI: https://doi.org/10.1016/j.dendro.2022.125959

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Published

26.07.2022

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How to Cite

Balzano, A., Čufar, K., Krže, L., & Merela, M. (2022). 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. Les/Wood, 71(1), 47-56. https://doi.org/10.26614/les-wood.2022.v71n01a05

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