Correlative imaging of collagen fibers and fibroblasts using CLEM optimized for picrosirius red staining and FIB/SEM tomography | Zendy

Shingo Hirashima | Zendy; Keisuke Ohta | Zendy; Tomonoshin Kanazawa | Zendy; Akinobu Togo | Zendy; Risa Tsuneyoshi | Zendy; Yoshihiro Miyazono | Zendy; Jingo Kusukawa | Zendy; Keiichiro Nakamura | Zendy

Open Access

Correlative imaging of collagen fibers and fibroblasts using CLEM optimized for picrosirius red staining and FIB/SEM tomography

Author(s) -

Shingo Hirashima,

Keisuke Ohta,

Tomonoshin Kanazawa,

Akinobu Togo,

Risa Tsuneyoshi,

Yoshihiro Miyazono,

Jingo Kusukawa,

Keiichiro Nakamura

Publication year - 2020

Publication title -

microscopy

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.545

H-Index - 52

eISSN - 2050-5701

pISSN - 2050-5698

DOI - 10.1093/jmicro/dfaa024

Subject(s) - staining , ultrastructure , materials science , biomedical engineering , negative stain , pathology , electron microscope , optics , medicine , physics

Conventional imaging for three-dimensional (3D) ultra-architectural analysis of collagen fibers and fibroblasts is time-consuming and requires numerous ultrathin sections to search the target area. Currently, no method allows 3D ultra-architectural analysis of predetermined areas including spatial relationships between collagen fibers and fibroblasts in vitro. Herein, we developed a new method for in vitro analysis of the 3D ultrastructure of fibroblasts and collagen fibers using CLEM optimized for picrosirius red staining and FIB/SEM tomography. Collagen fibers were observed between, rather than on top of, stacked cells. This method offers the advantage of mesoscopic and ultrastructural analysis, thus minimizing bias and ensuring accurate observation.

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