Lee T C, Myers E R, Hayes W C
Department of Anatomy, Royal College of Surgeons in Ireland, Dublin.
J Anat. 1998 Aug;193 ( Pt 2)(Pt 2):179-84. doi: 10.1046/j.1469-7580.1998.19320179.x.
En bloc staining with basic fuchsin is an established method for demonstrating microdamage in bone. Using transmitted light microscopy, variations in light intensity, depth of focus and magnification are necessary to distinguish fully-stained microcracks generated in vivo, from partially-stained or unstained artefactual cracks due to cutting and machining. This process is both difficult and time-consuming. In this study, 2 methods were used to examine fuchsin-stained microcracks in human rib sections, transmitted light and epifluorescence microscopy. No differences were found in crack number, density or length between the 2 methods indicating comparable accuracy. Using green epifluorescence, only microcracks containing fuchsin fluoresced orange against the darkfield background, enabling unstained, artefactual cracks to be screened out. Under UV epifluorescence, microcracks stained through the full 100 microm depth of the section fluoresced purple. Partially-stained artefactual cracks failed to fluoresce and were screened out. Epifluorescence is a simple, rapid and accurate screening method for differentiating fully-stained from artefactual microcracks in bone.
碱性品红整体染色是一种已确立的用于显示骨微损伤的方法。使用透射光显微镜时，为了将体内产生的完全染色的微裂纹与因切割和加工产生的部分染色或未染色的人为裂纹区分开来，需要调整光强度、焦深和放大倍数。这个过程既困难又耗时。在本研究中，使用了两种方法来检查人肋骨切片中品红染色的微裂纹，即透射光显微镜和落射荧光显微镜。两种方法在裂纹数量、密度或长度上均未发现差异，表明准确性相当。使用绿色落射荧光时，只有含品红的微裂纹在暗场背景下发出橙色荧光，从而能够筛选出未染色的人为裂纹。在紫外落射荧光下，贯穿切片全100微米深度染色的微裂纹发出紫色荧光。部分染色的人为裂纹不发荧光并被筛选出来。落射荧光是一种简单、快速且准确的筛选方法，用于区分骨中完全染色的微裂纹和人为微裂纹。
