不同固定剂和脱钙剂组合对大鼠和兔硬组织的影响，组织学处理指南。

The effect of different combinations of fixatives and decalcifying agents on rat and rabbit hard tissues, a guide for histologic processing.

机构信息

Dental Research Center, Dentistry Research Institute, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran.

Department of Oral and Maxillofacial Radiology, School of Dentistry, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran.

出版信息

Acta Histochem. 2022 Dec;124(8):151962. doi: 10.1016/j.acthis.2022.151962. Epub 2022 Oct 10.

DOI:10.1016/j.acthis.2022.151962

PMID:36228481

Abstract

BACKGROUND AND PURPOSE

In order to acquire the best method that can simultaneously maximize tissue morphology and staining quality, we compared the effect of different fixative and decalcifying solutions on the quality of rabbit and rat bone histology.

METHOD

Fifty-four rat hemimaxillae and 54 rabbit quarter-parietal bones were allocated into 3 fixation groups (formalin, 10 %sodium-phosphate-buffered-formalin and 10 %calcium-phosphate-buffered-formalin). Each fixative was divided into 6 groups and decalcified with 5 % and 10 % nitric acid (NA), 5 % and 10 % formic acid (FA), Gooding-Stewart liquid (GSL) and EDTA. Slide quality was evaluated on hematoxylin/eosin slides by 3 observers and mean-scores for total-cell-characteristics (TCC) and total-tissue-characteristics (TTC) were statistically analyzed.

RESULT

Significant differences in decalcification-time were observed in different combinations of decalcifiers and fixatives in both animals. In rats, TCC was better preserved when using 10 %NA/calcium-phosphate-buffered-formalin compared to 10 %NA/sodium-phosphate-buffered-formalin (P = 0.03). GSL/sodium-phosphate-buffered-formalin performed better than both other fixatives (P < 0.001). TCC differed among the decalcifiers in each of the fixatives. In rabbits, there were differences in TCC among the decalcifiers when formalin (P = 0.001) and sodium-phosphate-buffered-formalin (P = 0.01) were used. TTC only showed significant difference when 10 %FA was used in rats (P = 0.044), with formalin performing better than sodium-phosphate-buffered-formalin (P = 0.01).

CONCLUSION

Based on our results, if time is an issue, 10 %NA/calcium-phosphate-buffered-formalin could provide good cellular quality and if time is not a consideration, FA (5 % or 10 %) with sodium-phosphate-buffered-formalin followed by EDTA with formalin, would have the best performance. In rabbits, GSL provides the fastest results, regardless of the fixative and FA/sodium-phosphate-buffered-formalin gives the best cellular quality.

摘要

背景与目的

为了获得同时最大限度地提高组织形态和染色质量的最佳方法，我们比较了不同固定剂和脱钙剂对兔和鼠骨组织学质量的影响。

方法

54 个大鼠上颌骨半侧和 54 个兔额顶骨四分体被分配到 3 个固定组（福尔马林、10%磷酸缓冲福尔马林和 10%钙磷酸盐缓冲福尔马林）。每个固定剂分为 6 组，用 5%和 10%硝酸（NA）、5%和 10%甲酸（FA）、Gooding-Stewart 液（GSL）和 EDTA 脱钙。3 位观察者对苏木精/伊红载玻片上的切片质量进行评估，并对总细胞特征（TCC）和总组织特征（TTC）的总分进行统计学分析。

结果

在两种动物中，不同脱钙剂和固定剂组合的脱钙时间存在显著差异。在大鼠中，与 10%NA/钙磷酸盐缓冲福尔马林相比，10%NA/磷酸缓冲福尔马林的 TCC 保存更好（P=0.03）。GSL/磷酸缓冲福尔马林的效果优于其他两种固定剂（P<0.001）。在每种固定剂中，TCC 在脱钙剂之间存在差异。在兔中，福尔马林（P=0.001）和磷酸缓冲福尔马林（P=0.01）中 TCC 在脱钙剂之间存在差异。仅在大鼠中使用 10%FA 时 TTC 显示出显著差异（P=0.044），福尔马林的效果优于磷酸缓冲福尔马林（P=0.01）。

结论

根据我们的结果，如果时间是一个问题，10%NA/钙磷酸盐缓冲福尔马林可以提供良好的细胞质量，如果时间不是问题，5%或 10%FA 与磷酸缓冲福尔马林一起使用，然后用 EDTA 与福尔马林一起使用，效果最佳。在兔中，GSL 无论固定剂如何，都能提供最快的结果，FA/磷酸缓冲福尔马林能提供最好的细胞质量。