Patterns of hyaluronan staining are modified by fixation techniques
Patterns of hyaluronan staining are modified by fixation techniques
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DOI：
10.1177/002215549704500813
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发表时间：
1997-08-01
影响因子：
3.2
通讯作者：
Stern, R
中科院分区：
文献类型：
--
作者：
Lin, WQ;Shuster, S;Stern, R
组织切片透明质酸（HA）染色的表观强度随固定技术的不同而变化。我们使用从牛鼻软骨中提取的HA特异性结合肽检测了HA在正常人皮肤中的组织化学分布。透明质酸，特别是真皮中的透明质酸，在10%的酸性福尔马林和70%的乙醇中固定的切片中保存得最好。相比之下，常规10%中性缓冲福尔马林固定的切片HA染色强度要弱得多。此外，酸性福尔马林/乙醇固定切片在生理盐水孵育后保留了大部分表观透明质酸，而中性福尔马林固定切片中大部分可染色的透明质酸丢失。在用阿利新蓝染色的玻片中没有观察到这种明显的染色强度差异，这种染色方法被认为是对透明质酸和其他糖胺聚糖进行染色的。当切片首次在透明质酸酶中预孵育时，使用HA结合肽的染色完全不存在，而类似的阿利新蓝染色切片保留了大部分的染色强度。在使用透明质酸结合肽评估组织中透明质酸的分布时应谨慎，特别是在比较几个实验室的不同固定技术时。此外，使用阿利新蓝染色评价组织中HA含量的能力应该重新考虑。“基质”的磺化糖胺聚糖似乎是阿利新蓝的主要底物。
The apparent intensity of hyaluronan (HA) staining in tissue sections can vary as a function of fixation techniques. We examined the histochemical distribution of HA in normal human skin using an HA-specific binding peptide derived from bovine nasal cartilage. The HA, particularly in the dermis, was best preserved in sections fixed in 10% acid-formalin with 70% ethanol. In contrast, sections fixed in the routine 10% neutral-buffered formalin had a much weaker intensity of HA staining. Furthermore, acid-formalin/ethanol-fixed sections retained much of their apparent HA after incubation with saline, in contrast to the neutral formalin-fixed sections, in which most of the stainable HA was lost. Such marked differences in staining intensity were not observed in slides stained with Alcian blue, a procedure pressumed to stain HA as well as other glycosaminoglycans. Staining using the HA binding peptide was entirely absent when sections were first preincubated in hyaluronidase, whereas similar Alcian blue-stained sections retained most of their staining intensity. Caution should be exercised in evaluating the distribution of HA in tissues using the HA binding peptide, particularly when different fixation techniques among several laboratories are being compared. In addition, the ability to evaluate the HA content of tissues using Alcian blue staining should be reconsidered. The sulfated glycosaminolglycans of the ''ground substance'' appear to be the predominant substrates for Alcian blue.