Scanning electron microscopy (SEM) protocols for problematic plant, oomycete, and fungal samplesVise andre og tillknytning
2017 (engelsk)Inngår i: Journal of Visualized Experiments, E-ISSN 1940-087X, Vol. 2017, nr 120, artikkel-id e55031Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]
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.
sted, utgiver, år, opplag, sider
Journal of Visualized Experiments , 2017. Vol. 2017, nr 120, artikkel-id e55031
Emneord [en]
Agaricales, Critical point dryer, Cysts, Formaldehyde, Glutaraldehyde, Issue 120, Phellorinia, Plant biology, Plant development, Saprolegnia, Sputter coater
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-208129DOI: 10.3791/55031ISI: 000397847700036PubMedID: 28190042Scopus ID: 2-s2.0-85014381669OAI: oai:DiVA.org:kth-208129DiVA, id: diva2:1106127
Forskningsfinansiär
EU, Horizon 2020, 634429
Merknad
QC 20170607
2017-06-072017-06-072024-03-15bibliografisk kontrollert