Dolber P C, Spach M S
Department of Pediatrics, Duke University Medical Center, Durham, North Carolina 27710.
J Histochem Cytochem. 1993 Mar;41(3):465-9. doi: 10.1177/41.3.7679127.
The arrangement of collagen fibers has previously been studied with picrosirius red (PSR) staining and brightfield microscopy. We discovered that PSR staining can also be visualized by fluorescence microscopy. PSR-stained collagen was strongly fluorescent using excitation and barrier filters for rhodamine, and distracting background cytoplasmic fluorescence was drastically reduced with phosphomolybdic acid (PMA) treatment before PSR staining. The PMA-PSR fluorescence method was more sensitive than the brightfield PSR or PMA-PSR method, and permitted confocal microscopic study. We applied the method to the study of collagen fiber three-dimensional arrangement in perimysial and endomysial septa of the heart, showing the three-dimensional course of the fibers in stereo views generated by confocal microscopy. The PMA-PSR fluorescence method should be generally useful for accurately determining collagen fiber three-dimensional arrangement, a necessary prelude to mechanical modeling of collagen-reinforced tissues.
先前已通过天狼星红苦味酸（PSR）染色和明场显微镜对胶原纤维的排列进行了研究。我们发现，PSR染色也可以通过荧光显微镜观察到。使用罗丹明的激发和阻挡滤光片时，PSR染色的胶原具有强烈的荧光，并且在PSR染色前用磷钼酸（PMA）处理可大幅减少干扰性的背景细胞质荧光。PMA-PSR荧光法比明场PSR法或PMA-PSR法更灵敏，并且可用于共聚焦显微镜研究。我们将该方法应用于心脏肌束膜和肌内膜间隔中胶原纤维三维排列的研究，在共聚焦显微镜生成的立体视图中显示了纤维的三维走向。PMA-PSR荧光法通常应有助于准确确定胶原纤维的三维排列，这是对胶原增强组织进行力学建模的必要前奏。
