Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F23%3A00574918" target="_blank" >RIV/60077344:_____/23:00574918 - isvavai.cz</a>
Alternative codes found
RIV/60076658:12310/23:43907205
Result on the web
<a href="https://frontiersinzoology.biomedcentral.com/counter/pdf/10.1186/s12983-023-00507-x.pdf" target="_blank" >https://frontiersinzoology.biomedcentral.com/counter/pdf/10.1186/s12983-023-00507-x.pdf</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1186/s12983-023-00507-x" target="_blank" >10.1186/s12983-023-00507-x</a>
Alternative languages
Result language
angličtina
Original language name
Minimal resin embedding of SBF-SEM samples reduces charging and facilitates finding a surface-linked region of interest
Original language description
Background: For decoding the mechanism of how cells and organs function information on their ultrastructure is essential. High-resolution 3D imaging has revolutionized morphology. Serial block face scanning electron microscopy (SBF-SEM) offers non-laborious, automated imaging in 3D of up to ~ 1 mm3 large biological objects at nanometer-scale resolution. For many samples there are obstacles. Quality imaging is often hampered by charging effects, which originate in the nonconductive resin used for embedding. Especially, if the imaged region of interest (ROI) includes the surface of the sample and neighbours the empty resin, which insulates the object. This extra resin also obscures the sample’s morphology, thus making navigation to the ROI difficult.nResults: Using the example of small arthropods and a fish roe we describe a workflow to prepare samples for SBF-SEM using the minimal resin (MR) embedding method. We show that for imaging of surface structures this simple approach conveniently tackles and solves both of the two major problems—charging and ROI localization—that complicate imaging of SBF-SEM samples embedded in an excess of overlying resin. As the surface ROI is not masked by the resin, samples can be precisely trimmed before they are placed into the imaging chamber. The initial approaching step is fast and easy. No extra trimming inside the microscope is necessary. Importantly, charging is absent or greatly reduced meaning that imaging can be accomplished under good vacuum conditions, typically at the optimal high vacuum. This leads to better resolution, better signal to noise ratio, and faster image acquisition.nConclusions: In MR embedded samples charging is minimized and ROI easily targeted. MR embedding does not require any special equipment or skills. It saves effort, microscope time and eventually leads to high quality data. Studies on surface-linked ROIs, or any samples normally surrounded by the excess of resin, would benefit from adopting the technique.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10605 - Developmental biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2023
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Frontiers in Zoology
ISSN
1742-9994
e-ISSN
1742-9994
Volume of the periodical
20
Issue of the periodical within the volume
1
Country of publishing house
GB - UNITED KINGDOM
Number of pages
13
Pages from-to
29
UT code for WoS article
001057707300001
EID of the result in the Scopus database
2-s2.0-85168878106