Immunocytochemistry methods

Bendayan, M. (1984) Protein A-gold electron microscopic immunocytochemistry methods, applications, and limitations. J. Electron Micros. Tech. 1, 243-270. 

Morphological approaches in biomedical research can include a wide range of microscopes, but today typically employ immunocytochemistry that can give us information about individual liver cells containing the specific enzyme. Immunocytochemistry uses antibodies to bind proteins and labels to show protein s location. If, for example, the enzyme is a marker for inflammation, then the location of cells with this enzyme tells us which cell types have the inflammatory response. Thus, immunocytochemistry methods are broadly defined as individual studies of single cells or cell groups. The resulting data tell us about location of the enzyme. 

For more experienced users, individual chapters can be used to guide a specific part of the immunocytochemistry method. For example, if the user needs to... 

Fig. 7.1 Direct immunocytochemistry. In the direct immunocytochemistry method, the 1° antibody binds its antigen and the label is directly attached to it...

To help evaluate possible labels and methods, use Table 9.1. Note that the number ratings in Table 9.1 are based on the author s experience and should only be used to compare methods within this table. Detection resolution is the ability to localize the label to the exact site of the primary antibody. The direct immunocytochemistry method does this and is rated as a detection resolution of 10 (Table 9.1, Fig. 9.1a, arrow). With detection methods that have lower detection resolution the number decreases (Table 9.1, Fig. 9.1a, numbers). 

McDonald K. 1994. Electron microscopy and EM immunocytochemistry. Methods Cell Biol. 44 411-444. 

Webster P. 1999. The production of cryosections through fixed and cryoprotected biological material and their use in immunocytochemistry. Methods Mol. Biol. 117 46-76. 

Nassel DR, Ekstrom P (1997) Detection of neuropeptides by immunocytochemistry. Methods Mol Biol 72 71-101... 

Immunocytochemical methods have been widely applied to visualize proteins, carbohydrates, or lipids in sectioned material. The advantage of using immunocytochemistry is to be able to localize the molecules of interest within the tissue. Several procedures have been described. Basically, these procedures can be split into four main steps that are described in subheadings (1) tissue preparation, (2) the primary antibodies, (3) the visualization of the target, and (4) enhancement of signals with antibody complexes. In addition, a protocol for alkaline phosphatase will be presented in detail in Subheading 5. The terms primary and secondary antibodies refer to the order in which they are applied to the target. The immunocytochemical procedures are not limited to sectioned... 

Singh MB, Taylor PE, Knox RB. Special preparation methods for immunocyto-chemistry of plant cells, in Immunocytochemistry, A Practical Approach (Beesley JE, ed.), Oxford University Press, Oxford, UK, 1993, pp. 77-100. 

De Mey J, Moeremans M. Raising and testing polyclonal antibodies for immunocytochemistry, in Immunocytochemistry, Modern Methods and Applications (Polack JM, Van Noorden S, eds.), Wright, Bristol, UK, 1986, pp. 3-12. 

Sawada H, Esaki M. A simple flat embedding method for the correlative light and electron microscopic immunocytochemistry. J Electron Microsc 1998 47 535-537. 

Plant Cells and Tissues Structure-Function Relationships. Methods for the Cytochemical/Histochemical Localization of Plant Cell/Tissue Chemicals. Methods in Light Microscope Radioautography. Some Fluorescence Microscopical Methods for Use with Algal, Fungal, and Plant Cells. Fluorescence Microscopy of Aniline Blue Stained Pistils. A Short Introduction to Immunocytochemistry and a Protocol for Immunovi-sualization of Proteins with Alkaline Phosphatase. The Fixation of Chemical Forms on Nitrocellulose Membranes. Dark-Field Microscopy and Its Application to Pollen Tube Culture. Computer-Assisted Microphotometry. Isolation and Characterization of... 

Stemberger LA (1986) Immunocytochemistry, 3rd Edn. John Wiley and Sons, New York Van Noorden CJF, Frederiks WM (1993) Enzyme Histochemistry A Laboratory Manual of Current Methods. Bios Scientific Publishers Ltd., Oxford... 

Pirici D, Mogoanta L, Kumar Singh S, Pirici I, Margaritescu C, Simionescu C, Stanescu R (2009) Antibody elution method for multiple immunohistochemistry on primary antibodies raised in the same species and of the same subtype. J Histochem Cytochem 57(6) 567 575 Polak JM, Van Noorden S (1997) Introduction to immunocytochemistry. BIOS Scientific, Oxford... 

Tramu G, Pillez A, Leonardelli J (1978) An efficient method of antibody elution for the successive or simultaneous localization of two antigens by immunocytochemistry. J Histochem Cytochem 26 322 324... 

VanNoorden, S. (1986) Tissue preparation and immunostaining techniques for light microscopy, in Immunocytochemistry Modern Methods and Applications (Polak, J. M. and VanNoorden, S., eds.), Wright, Bristol, England, pp. 26-53. 

Gnstafsson, M. K. (1991) Skin the tapeworms before you stain their nervous system A new method for whole-mount immunocytochemistry. Parasitology Res. 77, 509-516. 

---