Abstract
AIMS--To develop and immunohistochemical staining method for cryostat and paraffin wax sections so that two different antigens in the same section of tissues could be detected by combining immunoenzyme and immunofluorescence techniques. METHODS--This double immunohistochemical staining method combines alkaline phosphatase-anti-alkaline phosphatase (APAAP) using New Fuchsin as a chromogen and indirect immunofluorescence. RESULTS--APAAP staining for one antigen of this double immunohistochemical staining method was observed under bright field conditions alternating with immunofluorescence for another antigen under ultraviolet light. The double exposed photograph of both easily identified the two signals within the same cell. CONCLUSIONS--This double immunohistochemical staining method can overcome the disadvantages of any masking effect of the double immunoenzymatic methods and the background problems of double immunofluorescence method especially when applied to paraffin wax sections. It also permits good morphological identification of the doubly stained cells which may be of crucial importance in studies on pathology specimens.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bologna M., Allen R., Dulbecco R. A method for double immunofluorescent staining by the indirect procedure with antibodies of the same isotype. J Immunol Methods. 1986 Jan 22;86(1):151–153. doi: 10.1016/0022-1759(86)90279-6. [DOI] [PubMed] [Google Scholar]
- Cordell J. L., Falini B., Erber W. N., Ghosh A. K., Abdulaziz Z., MacDonald S., Pulford K. A., Stein H., Mason D. Y. Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem. 1984 Feb;32(2):219–229. doi: 10.1177/32.2.6198355. [DOI] [PubMed] [Google Scholar]
- Falini B., Abdulaziz Z., Gerdes J., Canino S., Ciani C., Cordell J. L., Knight P. M., Stein H., Grignani F., Martelli M. F. Description of a sequential staining procedure for double immunoenzymatic staining of pairs of antigens using monoclonal antibodies. J Immunol Methods. 1986 Nov 6;93(2):265–273. doi: 10.1016/0022-1759(86)90199-7. [DOI] [PubMed] [Google Scholar]
- Lechago J., Sun N. C., Weinstein W. M. Simultaneous visualization of two antigens in the same tissue section by combining immunoperoxidase with immunofluorescence techniques. J Histochem Cytochem. 1979 Sep;27(9):1221–1225. doi: 10.1177/27.9.113453. [DOI] [PubMed] [Google Scholar]
- Murdoch A., Jenkinson E. J., Johnson G. D., Owen J. J. Alkaline phosphatase-fast red, a new fluorescent label. Application in double labelling for cell cycle analysis. J Immunol Methods. 1990 Aug 28;132(1):45–49. doi: 10.1016/0022-1759(90)90396-d. [DOI] [PubMed] [Google Scholar]
- Ramshaw A. L., Parums D. V. Combined immunohistochemical and immunofluorescence method to determine the phenotype of proliferating cell populations. J Clin Pathol. 1992 Nov;45(11):1015–1017. doi: 10.1136/jcp.45.11.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wagner L., Worman C. P. Color-contrast staining of two different lymphocyte subpopulations: a two-color modification of alkaline phosphatase monoclonal anti-alkaline phosphatase complex technique. Stain Technol. 1988 May;63(3):129–136. doi: 10.3109/10520298809107172. [DOI] [PubMed] [Google Scholar]