[Recent advances in molecular histochemical techniques].
[Recent advances in molecular histochemical techniques].
复制标题
分子组织化学技术的最新进展。
DOI：
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发表时间：
1999
期刊：
影响因子：
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通讯作者：
T. Koji
中科院分区：
文献类型：
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作者：
T. Koji
为了更好地理解细胞和组织的生理状态，有必要分析单个细胞中细胞类型或组织类型特异性基因表达的调控机制。因此，原位杂交（ISH）被认为是一种有效的分子组织化学方法。在过去的十年中，我们做了大量的努力，建立非放射性的ISH方案与胸腺嘧啶-胸腺嘧啶二聚或地高辛标记的合成寡核苷酸探针，包括广泛的优化预处理，可杂交RNA的评价和新的试验信号放大。此外，我们最初开发了一种新的方法，西南组织化学，它定位特定的转录调控因子，使我们能够了解ISH检测到的mRNA水平的波动在单个细胞中的基因的转录调控水平。在这里，我将回顾这些方法的概述以及最近的进展，并将看看我们可以通过组合它们看到什么。
To understand better the physiological states of cells and tissues, it is essential to analyze the regulatory mechanism of cell-type or tissue-type specific gene expression in individual cells. For this purpose, in situ hybridization (ISH) has been regarded as a powerful molecular histochemical method. Over the past decade, we made lots of efforts to establish nonradioactive ISH protocol with thymine-thymine dimerized or digoxigenin-labeled synthetic oligo-DNA probes, including extensive optimization of pretreatment, evaluation of hybridizable RNA and new trials of signal amplification. Furthermore, we have originally developed a new method, Southwestern histochemistry, which localizes specific transcription regulatory factors and allows us to understand the fluctuation of mRNA levels detected by ISH at the level of transcription regulation of the gene in individual cells. Here, I will review the outlines of these methods with recent progress and will see what we can see by combining them.