ABSTRACT
The renal glomerulus produces primary urine from blood plasma by ultrafiltration. The ultrastructure of the glomerulus is closely related to filtration function and disease development. The ultrastructure of glomeruli has mainly been evaluated using transmission electron microscopy; however, the volume that can be observed using transmission electron microscopy is extremely limited relative to the total volume of the glomerulus. Consequently, observing structures that exist in only one location in each glomerulus, such as the vascular pole, and evaluating low-density or localized lesions are challenging tasks. Array tomography (AT) is a technique used to analyze the ultrastructure of tissues and cells via scanning electron microscopy of serial sections. In this study, we present an AT workflow that is optimized for observing complete serial sections of the whole glomerulus, and we share several analytical examples that use the optimized AT workflow, demonstrating the usefulness of this approach. Overall, this AT workflow can be a powerful tool for structural and pathological evaluation of the glomerulus. This workflow is also expected to provide new insights into the ultrastructure of the glomerulus and its constituent cells.
Footnotes
Author contributions
Conceptualization: T.M., K.I.; Methodology: T.M., N.H., Y.K., M.K., J.Y., S.K., T. Shindo, M.S., J.A.O.T., T.O., T.N.-K., M.I., K.I.; Investigation: T.M., K.I.; Resources: N.H., M.T., K.A., S.I., K.K., S.M., M.A., S.H., K.O., Y.S., K.I.; Data curation: T.M., N.H., Y.K., M.K.; Writing - original draft: T.M., K.I.; Writing - review & editing: T.M., K.I.; Visualization: T.M., N.H., Y.K., M.K., M.S., H.K.; Supervision: S.S., T. Sakai, Y.S.; Project administration: T.M., K.I.; Funding acquisition: T.M., J.A.O.T., K.I.
Funding
This work was supported in part by the Japan Society for the Promotion of Science (JSPS) KAKENHI (grants 22J12056 and 23K14433 to T.M.; 22K08363 to J.A.O.T.; 17K08521 and 22K06799 to K.I.) and the Foundation of Strategic Research Projects in Private Universities from the Ministry of Education, Culture, Sports, Science and Technology of Japan (grant S1311011; Juntendo University).
Data availability
All relevant data can be found within the article and its supplementary information.
Special Issue
This article is part of the Special Issue ‘Imaging Cell Architecture and Dynamics’, guest edited by Lucy Collinson and Guillaume Jacquemet. See related articles at https://journals.biologists.com/jcs/issue/137/20.