Correlative volume-imaging using combined array tomography and FIB-SEM tomography with beam deceleration for 3D architecture visualization in tissue

Abstract Focused ion beamed (FIB) SEM has a higher spatial resolution than other volume-imaging methods owing to the use of ion beams. However, in this method, it is challenging to analyse entire biological structures buried deep in the resin block. We developed a novel volume-imaging method by comb...

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Bibliographic Details
Published in:Microscopy 2022-06, Vol.71 (3), p.187-192
Main Authors: Hirashima, Shingo, Ohta, Keisuke, Rikimaru-Nishi, Yukiko, Togo, Akinobu, Funatsu, Takashi, Tsuneyoshi, Risa, Shima, Yuichi, Nakamura, Kei-ichiro
Format: Article
Language:English
Subjects:
Deceleration
Imaging, Three-Dimensional - methods
Microscopy, Electron, Scanning
Tomography
Tomography, X-Ray Computed
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Summary:Abstract Focused ion beamed (FIB) SEM has a higher spatial resolution than other volume-imaging methods owing to the use of ion beams. However, in this method, it is challenging to analyse entire biological structures buried deep in the resin block. We developed a novel volume-imaging method by combining array tomography and FIB-SEM tomography and investigated the chondrocyte ultrastructure. Our method imparts certainty in determining the analysis area such that cracks or areas with poor staining within the block are avoided. The chondrocyte surface showed fine dendritic processes that were thinner than ultrathin sections. Upon combination with immunostaining, this method holds promise for analysing mesoscopic architectures.
ISSN:2050-5698
2050-5701
DOI:10.1093/jmicro/dfac015