Immunohistochemical imaging

Fig. 1. Changes in immunoreactivity for the 72 subunit of the GABAa-R in the rat hippocampus during pregnancy and after delivery. (A, B) Representative immunohistochemical images of the distribution of the 72 subunit in the dentate gyrus (A) and in the CA1 region (B) in hippocampal sections from rats in estrus (E) at day 15 (P15), 19 (PI 9), or 21 (P21) of pregnancy or at 2 (pp2) or 7...

Immunohistochemical imaging Eu3+-W1024, Tb3+-W14016 (Wallac/PerkinElmer) Siivola et al. (2000)... 

Immunohistochemical staining is very different from routine chromogenic stains such as H E. To utilize IHC stains for quantitative purposes, the user must understand these differences and perform the stain in both a standardized and repeatable manner. As has been described, image analysis... 

Optical imaging methods for hypoxia are not yet applicable in vivo but immunohistochemical antibody detection of nitroimi-dazoles (e.g. pimonidazole) has been used extensively for in vitro work and biopsy tissue staining (89). 

In this chapter we report recent analytical applications of CL imaging for the detection of biospecific reactions in macrosamples such as microtiter plates of different format (96 or 384 wells), filter membranes and irregular surfaces represented by specimens related to the cultural heritage, and results obtained when the CCD detector is coupled with optical microscopy for enzyme localization, immunohistochemical reactions, and complementary DNA (cDNA) detection (Table 1). 

M. Ogawa, S. Ishino, T. Mukai, D. Asano, N. Teramoto, H. Watabe, N. Kudomi, M. Shiomi, Y. Magata, H. lida, H. Saji, F-FDG accumulation in atherosclerotic plaques Immunohistochemical and PET imaging study, J. Nucl. Med. 45 (2004) 1245-1250. 

A related approach was used by Ranefall et al. (1998) to quantify images of immunohistochemically stained cell nuclear Ki-67 antigen and cyclin A protein in bladder carcinoma tissue. They combined automatic, computerized image analysis with appropriate controls and reference material. This approach is superior to the automatic method without... 

The image of the control cells serves as a standard control regarding image qualities such as illumination and color properties. Since control cells possess known characteristics regarding antigen expression to be examined in tissue sections, they serve as a means to control and standardize immunohistochemical data (Ranefall et al., 1998). 

Esteban, J. M., Ahn, C., Mehta, P., and Battifora, P. 1994b. Biologic significance of quantitative estrogen receptor immunohistochemical assay by image analysis in breast cancer. Am. J. Pathol. 102 158c. 

Hatanaka, Y., Hashizume, K., Kamihara, Y., Itoh, H., Tsuda, H., Osamura, R. Y., and Tany, Y. 2001. Quantitative immunohistochemical evaluation of HER2/neu expression with Herceptest in breast carcinoma by image analysis. Pathol. Int. 51 33-36. 

James, J. D., and Hauer-Jensen, M. 1999. Effects of fixative and fixation time for quantitative computerized image analysis of immunohistochemical staining. J. Histotechnol. 22 109-111. 

Kim, D., Gregory, C. W., Smith, G. J., and Mohler, J. L. 1999a. Immunohistochemical quantitation of androgen receptor expression using color video image analysis. Cytometry 35 2-10. 

Mangham, D. C., and Isaacson, P. G. 1999. A novel immunohistochemical detection system using mirror image complementary antibodies (MICA). Histopathology 35 129-133. 

Additionally, the principle of garbage-in, garbage-out cannot be overstated. There is no point in attempting to quantify IHC slides unless one is sure that the immunohistochemical procedure has been produced in a reproducible fashion. Automation and standardization in the process of preparing the IHC slides are very important components for successful image analysis. 

It is possible to predict the effect of treatment in cancer in a noninvasive manner by apoptosis imaging in vivo after radiotherapy by using I-Annexin V. Brain tumors were implanted in susceptible (nude) mice. By 6 h after receiving levels of x-irradiation as low as 2 Gy both autoradiography and immunohistochemical staining showed more apoptosis in the tumors of irradiated groups than in the control group (Watanabe et al, 2006). 

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