Pradhan Prita, Rajesh Nachiappa Ganesh, Badhe Bhawana Ashok, Ilanchezian Kalaivizhi, Manimehalai Dharmalingam, Jyothish Arikkotil
Currently working in Department of Pathology, Kalinga Institute of Medical Sciences, Bhubaneswar, Odisha, India.
Department of Pathology, Jawaharlal Institute of Post graduate Medical Education and Research (JIPMER), Puducherry, India.
Indian J Pathol Microbiol. 2019 Jul-Sep;62(3):423-429. doi: 10.4103/IJPM.IJPM_853_18.
Utility of modified Hammersmith protocol in the deacalcification and/or softening of tissues and samples in a histopathology laboratory were studied. The object of the study was to prepare a novel method for softening/decalcifying tissue for histopathology.
All the hard tissues received in the histopathology section were received in 10% neutral buffered formalin and then placed in freshly prepared combination of 10 mL of concentrated formaldehyde and 5 mL of 10% formic acid in 85 mL distilled water was used for decalcification. The tissue was checked for evidence of adequate decalcification/softening every 6 hours. Those which were decalcified/softened were sent for routine tissue processing and staining, while those which were not, were again placed in formalin. The process was repeated until the tissue was ready for further processing. The routine sections of these slides were reviewed for morphology and stain quality along with special stains and immunohistochemistry performed. The time taken for decalcification, the variables most likely to affect decalcification, the morphology and staining characteristics were documented. Statistical analysis was done to determine the effect of softening/decalcification process on each variable.
A total of 201 blocks in 119 specimens from humans including 61 males and 58 females were studied. Time taken was found to have a significant correlation only with the nature of the tissue (bone vs nonbone) and not with any other parameter viz. age, gender, specimen size, type of bone, and nature of pathology.
This novel and modified method has circumvented the common problems of overdecalcification, preserved morphology, and produced consistent results without interfering with special stains and immunohistochemistry.
研究改良的哈默史密斯方案在组织病理学实验室中对组织和样本进行脱钙和/或软化的效用。本研究的目的是制备一种用于组织病理学的新型组织软化/脱钙方法。
组织病理学切片接收的所有硬组织均置于10%中性缓冲福尔马林中，然后放入新鲜配制的脱钙剂中，该脱钙剂由10 mL浓甲醛、5 mL 10%甲酸和85 mL蒸馏水组成。每6小时检查一次组织是否有充分脱钙/软化的迹象。已脱钙/软化的组织送去进行常规组织处理和染色，未脱钙/软化的组织则再次放入福尔马林中。重复该过程，直到组织准备好进行进一步处理。对这些玻片的常规切片进行形态学和染色质量评估，并进行特殊染色和免疫组织化学检测。记录脱钙所需时间、最可能影响脱钙的变量、形态学和染色特征。进行统计分析以确定软化/脱钙过程对每个变量的影响。
共研究了119例人类标本中的201个组织块，其中男性61例，女性58例。发现脱钙时间仅与组织性质（骨组织与非骨组织）显著相关，与年龄、性别、标本大小、骨类型和病理性质等其他参数无关。
这种新颖的改良方法避免了过度脱钙的常见问题，保留了形态学，并在不干扰特殊染色和免疫组织化学的情况下产生了一致的结果。
