Immunohistochemical Double Staining with Immunogold-Silver and Alkaline Phosphatase to Identify Nuclear Markers of Cellular Proliferation
作者:
ShibuyaMakoto,
ItoSatoyuki,
DavisRichard L,
HoshinoTakao,
期刊:
Biotechnic&Histochemistry
(Taylor Available online 1993)
卷期:
Volume 68,
issue 1
页码: 17-19
ISSN:1052-0295
年代: 1993
DOI:10.3109/10520299309105571
出版商: Taylor&Francis
关键词: Immunohistochernistry;double staining;immunogold-silver staining;alkaline phosphatase staining;bromodeoxyuridine;iododeoxyuridine;cell kinetics
数据来源: Taylor
摘要:
To facilitate cell kinetics studies of brain tumors labeled with thymidine analogs, we developed a new method to identify nuclei labeled sequentially with bromodeoxyuridine (BUdR) and iododeoxyuridine (IUdR) by double staining with immunogold-silver and alkaline phosphatase. Patients received an intraoperative infusion of BUdR: excised tumor specimens were immediately labeled with IUdR in vitro. fixed with 70% alcohol, embedded in paraffin, and cut into 6pmsections. The sections were incubated first with BR-3. a monoclonal antibody that recognizes only BUdR, and then with IU-4. a monoclonal antibody that recognizes both BUdR and IUdR: sections were counterstained with hematoxylin to identify unlabeled nuclei. Nuclei labeled only with IUdR stained red, whereas those labeled with BUdR or with both BUdR and IUdR stained black against a red background: unlabeled nuclei stained blue. This method was the most efficient differential staining technique to identify nuclei labeled only with IUdR and those labeled with BUdR among unlabeled nuclei.
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