Zakout Yosef Mohamed Azzam, Salih Magdi M, Ahmed H G
Department of Histopathology and Cytology, Faculty of Medical Laboratory Sciences, University of Khartoum, Khartoum, Sudan.
Biotech Histochem. 2010 Apr;85(2):93-8. doi: 10.3109/10520290903126883.
Glycogen is demonstrated in a number of lesions and is diagnostically significant, particularly in certain tumors. To stain glycogen accurately, it is essential to choose a suitable fixative, temperature and staining method. We used rabbit liver to assess these variables. Specimens were fixed in three fixatives at two temperatures: 10% formalin, neutral buffered formalin (NBF) and Bouin's solution at 37 and 4 degrees C. Seventy-two paraffin sections were prepared and stained with periodic acid-Schiff (PAS), hexamine (methenamine) silver and Best's carmine methods. Negative control sections using diastase digestion were used for all methods to confirm the presence of glycogen. For the PAS reaction, Bouin's fixative gave better results at both temperatures compared to the other fixatives. For hexamine (methenamine) silver, the quality of staining was improved for tissues fixed in both 10% formalin and NBF at 37 degrees C compared to Bouin's solution. Both 10% formalin and NBF at 4 degrees C gave better results than Bouin's solution. For Best's carmine, Bouin's solution gave the best results for tissues fixed at 4 degrees C. Fixation of tissues with NBF at 37 degrees C gave the best quality staining. We concluded that the quality of glycogen staining in paraffin sections is greatly affected by both the fixative and the temperature of fixation.
糖原在许多病变中都有显示，且具有诊断意义，尤其是在某些肿瘤中。为了准确地对糖原进行染色，选择合适的固定剂、温度和染色方法至关重要。我们使用兔肝来评估这些变量。标本在两种温度下用三种固定剂固定：10%福尔马林、中性缓冲福尔马林（NBF）和波因氏液，温度分别为37℃和4℃。制备了72张石蜡切片，并用过碘酸-希夫（PAS）、六胺（乌洛托品）银和贝斯特胭脂红方法进行染色。所有方法均使用淀粉酶消化的阴性对照切片来确认糖原的存在。对于PAS反应，与其他固定剂相比，波因氏固定剂在两种温度下都能产生更好的结果。对于六胺（乌洛托品）银染色，与波因氏液相比，在37℃下用10%福尔马林和NBF固定的组织染色质量有所提高。4℃下的10%福尔马林和NBF都比波因氏液产生更好的结果。对于贝斯特胭脂红，波因氏液对4℃固定的组织效果最佳。NBF在37℃下固定组织能获得最佳质量的染色。我们得出结论，石蜡切片中糖原染色的质量受固定剂和固定温度的影响很大。
