使用针对 picrosirius 红染色和 FIB/SEM 断层扫描进行优化的 CLEM 对胶原纤维和成纤维细胞进行相关成像。

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

机构信息

Division of Microscopic and Developmental Anatomy, Department of Anatomy, Kurume University School of Medicine, 67 Asahi-machi, Kurume, Fukuoka 830-0011, Japan.

Dental and Oral Medical Center, Kurume University School of Medicine, 67 Asahi-machi, Kurume, Fukuoka 830-0011, Japan.

出版信息

Microscopy (Oxf). 2020 Oct 30;69(5):324-329. doi: 10.1093/jmicro/dfaa024.

DOI:10.1093/jmicro/dfaa024

PMID:32406508

Abstract

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.

摘要

传统的用于三维（3D）胶原纤维和成纤维细胞超微结构分析的成像方法既耗时又需要大量超薄切片来搜索目标区域。目前，尚无方法可对体外预定区域的 3D 超微结构进行分析，包括胶原纤维和成纤维细胞之间的空间关系。在此，我们开发了一种新的方法，使用针对 picrosirius 红染色和 FIB/SEM 断层扫描进行了优化的 CLEM，对体外成纤维细胞和胶原纤维的 3D 超微结构进行分析。胶原纤维观察到在堆叠细胞之间，而不是在堆叠细胞之上。该方法具有介观和超微结构分析的优势，从而最大程度地减少了偏差并确保了准确的观察。