Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest
Serial block-face scanning electron microscopy (SBF-SEM or SBEM) is a state-of-the-art groundbreaking technique that offers volume imaging of ~1 mm3 large biological objects at nanometer-scale resolution. We have successfully applied the minimal resin (MR) embedding method to the sample preparation protocol and showed that this simple approach tackles simultaneously two major problems of SBF-SEM: charging and finding the region of interest (ROI).
Volume electron microscopy (for 3D reconstructions), to which SBF-SEM belongs, was named by the journal Nature as one of the “Seven technologies to watch in 2023” alongside the James Webb Space Telescope, CRISPR, and others. SBF-SEM benefits from being highly automated and offering to obtain stacks of high-resolution images from “large volumes”. Hence it enables studies on cell organelles and their compartments in whole bodies of small invertebrates, such as insects. Frequent sample charging lowers resolution and may completely disrupt imaging. Preparing samples using the MR embedding minimizes charging and considerably up-levels the final image quality. Additionally, amount of time and workload is saved, since the ROI can be more easily targeted in the microscope. We believe that our findings will make SBF-SEM more accessible, so that it can be easily incorporated into the toolkit of any biologist, including entomologists.
Our study was published in the BMC journal Frontiers in Zoology, a journal supported by the German Zoological Society.
This project received funding from Czech-BioImaging (funded by the Ministry of Education, Youth and Sports CR): project LM2018129 to Barbora Konopová. The data were collected at the Laboratory of electron microscopy (Czech-BioImaging Facility).
Konopová, B., Týč, J.: Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest. Front Zool 20, 29 (2023).
