Vigliani R
Unità Operativa di Anatomia e Istologia Patologica, Ospedale E. Agnelli, ASL 10, via Brigata Cagliari 39, 10064 Pinerolo, Torino.
Pathologica. 2003 Jun;95(3):140-5.
In the literature the usefulness of the Perls reaction for Iron (Fe+++) stores estimation on bone marrow is largely accepted. However, due to decalcification, there is a considerable disagreement about the accuracy of this staining on osteomedullary tissue. In fact loss of Fe+++ has been claimed by some Authors, and denied by others. In order to check this controversial point in this paper Perls reaction, as a methodological variant, was directly anticipated on fragments (pre-Perls). In this way numerous osteomedullary sternal specimens from autopsies (non hematological cases) were observed in comparison with normal Perls reaction on sections (standard-Perls). In standard-Perls series (A) decalcification was achieved by HCl (alone or in combination with EDTA) or by formic acid. On pre-Perls treated fragments decalcification was an intrinsic effect of the hydrochloride component during Prussian Blue formulation. Nine different procedures of fixation were tested respectively for standard-Perls and pre-Perls. Pre-Perls was performed on pre-fixed (B series) as well as post-fixed material (C series). For the microscopical evaluation was applied the classical grading system proposed by Lundin for histology. Maximum grade was only observed in B and C, was the same in both these series and varied from 2 to 4 depending on autopsic case. Results strongly indicate the superiority of pre-Perls versus standard-Perls, with a differential grading ranging from 2 to 4 grades, in seven procedures of B (two procedures were equivalent in A and B) and in all the procedures of C. The comparison between B and C series showed inferiority of B in four procedures. For control (D series) standard-Perls was systematically performed on sections from pre-Perls: no differences between pre-Perls and pre-Perls + standard-Perls were observed indicating a complete permeation of fragments by Perls reagents. In general terms morphology in pre-Perls B was better than in pre-Perls C and substantially identical to standard procedures of A. Other data are analytically reported.
Pre-Perls staining (with special reference to five procedures of B) seems to be recommendable for decalcified bone marrow biopsies, in conjunction with standard processing, when an high sensitivity in detection of Fe+++ deposits is required or in absence of aspirate. Pathologists could subsequently decide if this methodological variant is suitable or not for an effective alternative choice in their routine according to morphological and immunohistochemical adequacy.
在文献中，珀尔斯反应对于评估骨髓中铁（Fe+++）储存量的用途已被广泛认可。然而，由于脱钙处理，对于这种染色方法在骨骨髓组织上的准确性存在相当大的分歧。事实上，一些作者声称存在Fe+++的损失，而另一些作者则予以否认。为了验证这一有争议的问题，在本文中，作为一种方法学变体，珀尔斯反应被直接应用于碎片（预珀尔斯反应）。通过这种方式，观察了大量来自尸检（非血液学病例）的胸骨骨髓标本，并与切片上的正常珀尔斯反应（标准珀尔斯反应）进行了比较。在标准珀尔斯反应系列（A）中，脱钙通过盐酸（单独或与乙二胺四乙酸联合使用）或甲酸实现。在经过预珀尔斯反应处理的碎片上，脱钙是普鲁士蓝形成过程中盐酸成分的固有作用。分别对标准珀尔斯反应和预珀尔斯反应测试了九种不同的固定程序。预珀尔斯反应在预固定材料（B系列）以及后固定材料（C系列）上进行。对于显微镜评估，应用了伦丁提出的经典组织学分级系统。最高等级仅在B和C中观察到，这两个系列中的等级相同，并且根据尸检病例从2到4不等。结果强烈表明，在B系列的七个程序（A和B中有两个程序相当）以及C系列的所有程序中，预珀尔斯反应优于标准珀尔斯反应，分级差异为2到4级。B和C系列的比较显示，在四个程序中B较差。作为对照（D系列），在预珀尔斯反应的切片上系统地进行标准珀尔斯反应：未观察到预珀尔斯反应和预珀尔斯反应 + 标准珀尔斯反应之间的差异，表明珀尔斯试剂完全渗透到碎片中。一般来说，预珀尔斯反应B中的形态比预珀尔斯反应C中的更好，并且与A的标准程序基本相同。其他数据将进行分析报告。
当需要对Fe+++沉积物具有高检测灵敏度或没有吸出物时，结合标准处理，预珀尔斯染色（特别参考B系列的五个程序）似乎适用于脱钙骨髓活检。病理学家随后可以根据形态学和免疫组织化学的充分性决定这种方法学变体在其日常工作中是否适合作为有效的替代选择。
