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HOPE技术为病理学开辟了众多新的可能性。

The HOPE technique opens up a multitude of new possibilities in pathology.

作者信息

Vollmer E, Galle J, Lang D S, Loeschke S, Schultz H, Goldmann T

机构信息

Clinical & Experimental Pathology, Research Center Borstel, Germany.

出版信息

Rom J Morphol Embryol. 2006;47(1):15-9.

Abstract

Fixation of tissues with formalin results in well-preserved morphology but to a high degree leads to degradation of nucleic acids, which substantially constricts the spectrum of applicable molecular techniques. The novel HOPE-fixative with subsequent paraffin embedding, as an alternative to formalin, has been shown to result in a morphological preservation comparable to formalin-fixed, paraffin-embedded specimens. Due to a similar workflow like in formalin-fixation and paraffin embedding, the HOPE technique can be successfully established within any pathological institute. We have shown that DNA, RNA and proteins are protected in HOPE-fixed, paraffin-embedded tissues for at least eight years. Moreover, we described procedures which permit successful application of all common molecular techniques such as in situ hybridization targeting either DNA or RNA, immunohistochemistry without antigen retrieval and for formalin-refractory antigens, PCR, RT-PCR, Western blot, Northern blot, and transcription microarrays to HOPE-fixed, paraffin-embedded tissues. Furthermore, HOPE-fixed tissues can be used for the construction of tissue microarrays for enhanced high-throughput analyses on the molecular level. Using the HOPE technique as its crucial methodological base, ex vivo model systems could be established, e.g. for the simulation of early events in human infections and detection of chemotherapy resistances in human cancer. In addition to tissues, cell-culture preparations have been prepared utilizing the HOPE technique, which were then successfully applied to in situ hybridization targeting mRNA or immunocytochemistry with excellent preservation of morphological details. Taken together, the HOPE technique to date represents an alternative fixation that is, in contrary to other procedures, scientifically broadly analyzed. Therefore new possibilities are opened up especially within the rapidly growing field of molecular pathology.

摘要

用福尔马林固定组织可使形态保存良好,但在很大程度上会导致核酸降解,这极大地限制了适用分子技术的范围。新型的HOPE固定剂结合后续石蜡包埋作为福尔马林的替代方法,已被证明能产生与福尔马林固定、石蜡包埋标本相当的形态保存效果。由于其工作流程与福尔马林固定和石蜡包埋相似,HOPE技术可在任何病理机构成功建立。我们已经表明,在HOPE固定、石蜡包埋的组织中,DNA、RNA和蛋白质至少能被保存八年。此外,我们还描述了一些程序,这些程序允许将所有常见的分子技术,如针对DNA或RNA的原位杂交、无需抗原修复的免疫组织化学以及针对福尔马林难处理抗原的免疫组织化学、PCR、RT-PCR、蛋白质免疫印迹、RNA印迹和转录微阵列,成功应用于HOPE固定、石蜡包埋的组织。此外,HOPE固定的组织可用于构建组织微阵列,以增强分子水平上的高通量分析。以HOPE技术作为关键的方法学基础,可以建立体外模型系统,例如用于模拟人类感染的早期事件和检测人类癌症中的化疗耐药性。除了组织,还利用HOPE技术制备了细胞培养制剂,然后成功地将其应用于针对mRNA的原位杂交或免疫细胞化学,形态细节保存良好。综上所述,迄今为止,HOPE技术是一种替代固定方法,与其他方法不同,它在科学上得到了广泛分析。因此,特别是在快速发展的分子病理学领域开辟了新的可能性。

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