Electron microscopy (EM) is a key tool for studying the microstructure of wood; however, observing uncoated samples poses a challenge due to surface charging. This study aims to identify the critical voltage that allows for the effective observation of uncoated wood samples without significant loading. As part of the experiment, samples of different wood species were tested, including Acacia (Robinia pseudoacacia L.), Oak (Quercus robur L.), Maple (Acer pseudoplatanus spp.), Ash (Fraxinus excelsior L.), Spruce (Picea abies (L.) Karst.), Thermowood (Thermal modifed Spruce), Garapa (Apuleia leiocarpa), Ipé (Handroanthus spp.), Merbau (Intsia bijuga), and Massaranduba (Manilkara spp.). Several methods were tested for surface preparation for SEM analysis, including the use of a circular saw, a hand milling machine, and a microtome. The results show that the optimal voltage for observing uncoated wood samples varied depending on the wood species. Regarding the selection of wood species and the results obtained, it was found that uncoated samples could be effectively observed. This finding suggests that practical observations can be accelerated and more cost-effective, as all wood species exhibited the required voltage range of 1 kV to 1.6 kV. Additionally, it was determined that using a secondary electron detector was optimal for such observations, as it provided a sufficiently strong signal even at relatively low voltages. Conversely, when using a backscattered electron detector, it was more beneficial to use coated samples to achieve a sufficient signal at higher voltages. This study brings new knowledge that will facilitate further research and applications of electron microscopy in the study of other wood species or wood-based materials.

Biotechnical Faculty University of Ljubljana Jamnikarjeva 101 1000 Ljubljana Slovenia

Department of Engineering Sciences and Mathematics Wood Science and Engineering Luleå University of Technology Forskargatan 1 93187 Skellefteå Sweden

Department of Wood Processing and Biomaterials Faculty of Forestry and Wood Sciences Czech University of Life Sciences Prague Kamýcká 1176 16500 Prague Czech Republic

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