Scanning electron microscopy (SEM) protocols for problematic plant, oomycete, and fungal samples

M. Angélica Bello, Yolanda Ruiz-León, J. Vladimir Sandoval-Sierra, Svetlana Rezinciuc, Javier Diéguez-Uribeondo

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Common problems in the processing of biological samples for observations with the scanning electron microscope (SEM) include cell collapse, treatment of samples from wet microenvironments and cell destruction. Using young floral tissues, oomycete cysts, and fungi spores (Agaricalesas examples, specific protocols to process delicate samples are described here that overcome some of the main challenges in sample treatment for image capture under the SEM. Floral meristems fixed with FAA (Formalin-Acetic-Alcohol) and processed with the Critical Point Dryer (CPD) did not display collapsed cellular walls or distorted organs. These results are crucial for the reconstruction of floral development. A similar CPD-based treatment of samples from wet microenvironments, such as the glutaraldehyde-fixed oomycete cysts, is optimal to test the differential growth of diagnostic characteristics (e.g., the cyst spines) on different types of substrates. Destruction of nurse cells attached to fungi spores was avoided after rehydration, dehydration, and the CPD treatment, an important step for further functional studies of these cells. The protocols detailed here represent low-cost and rapid alternatives for the acquisition of good-quality images to reconstruct growth processes and to study diagnostic characteristics.

Original languageEnglish (US)
Article numbere55031
JournalJournal of Visualized Experiments
Volume2017
Issue number120
DOIs
StatePublished - Feb 3 2017

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Oomycetes
Fungi
Electron Scanning Microscopy
Cysts
Scanning electron microscopy
Glutaral
Spores
Dehydration
Formaldehyde
Acetic Acid
Image quality
Alcohols
Electron microscopes
Tissue
Scanning
Cellular Microenvironment
Meristem
Fluid Therapy
Substrates
Therapeutics

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Scanning electron microscopy (SEM) protocols for problematic plant, oomycete, and fungal samples. / Bello, M. Angélica; Ruiz-León, Yolanda; Sandoval-Sierra, J. Vladimir; Rezinciuc, Svetlana; Diéguez-Uribeondo, Javier.

In: Journal of Visualized Experiments, Vol. 2017, No. 120, e55031, 03.02.2017.

Research output: Contribution to journalArticle

Bello, M. Angélica ; Ruiz-León, Yolanda ; Sandoval-Sierra, J. Vladimir ; Rezinciuc, Svetlana ; Diéguez-Uribeondo, Javier. / Scanning electron microscopy (SEM) protocols for problematic plant, oomycete, and fungal samples. In: Journal of Visualized Experiments. 2017 ; Vol. 2017, No. 120.
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