Bobrow M N, Moen P T
NEN Life Science Products, Boston, Massachusetts, USA.
Curr Protoc Cytom. 2001 May;Chapter 8:Unit 8.9. doi: 10.1002/0471142956.cy0809s11.
This unit provides a series of techniques that dramatically improve the sensitivity of direct and indirect in situ detection. TSA is a peroxidase-based signal amplification system which is compatible with all in situ hybridization as well as immunocytochemical detection systems. The assay is performed on a glass slide and combines the use of fluorescent probes in either direct or indirect format using either an enzyme-labeled streptavidin or antibody. Protocols are provided for brightfield and fluorescent detection of single DNA targets, DNA or RNA in cultured cells, multitarget detection for DNA/RNA and DNA and RNA. In addition a protocol is included for the TSA plus system, a DNP-based detection system using horseradish peroxidase and alkaline phosphatase.
本单元提供了一系列能显著提高直接和间接原位检测灵敏度的技术。TSA是一种基于过氧化物酶的信号放大系统，它与所有原位杂交以及免疫细胞化学检测系统兼容。该检测在载玻片上进行，使用酶标记的链霉亲和素或抗体，以直接或间接形式结合荧光探针的使用。提供了用于单个DNA靶标、培养细胞中的DNA或RNA的明场和荧光检测、DNA/RNA和DNA与RNA的多靶标检测的方案。此外，还包括TSA plus系统的方案，这是一种基于DNP的检测系统，使用辣根过氧化物酶和碱性磷酸酶。
