Light microscopy immunocytochemistry

Cote SL, Ribero-da-silva A, Cuello AC. 1994. Current protocols for light microscopy immunocytochemistry. Immuno-histochemistry II, Cuello AC, editor. Chichester John Wiley Sons. 

C6t6, A., Ribeiro-Da-Silva, A., and Cuello, A. C. (1993) Current protocols for light microscopy immunocytochemistry, in Immunohistochemistry II. IBRO Handbook Series Methods in Neurosciences, vol. 14 (Cuello, A. C., ed.), Wiley, Chichester, UK, pp. 147-168. 

JacksonP, Blythe D. Immunolabeling techniques Lor light microscopy, in Immunocytochemistry, A Practical Approach (Beesley JE, ed.), Oxford University Press, Oxford, UK, 1993, pp. 15-41. 

In addition to the standard set of tissues specified in Table 7.8, observations during the course of the study or in other previous studies may dictate that additional tissues be collected or special examinations (e.g., special stains, polarized light or electron microscopy, immunocytochemistry, or quantitative morphometry) be undertaken to evaluate the relevance of, or understand the mechanisms underlying, certain observations. 

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. 

Experience has shown that the desired routine sizes are 5, 10, and 15 nm. All these are useful for transmission electron microscope studies and the 5 nm gold is recommended for light microscopy. Gold spheres of 30 nm are occasionally used for scanning electron microscope immunocytochemistry. A1 nm probe is available and might be useful for both light and electron microscopy after silver enhancement. 

Immunolabeling for electron microscopy is theoretically identical to immunolabeling for light microscopy. The problems discussed in Chapter 17 describing light microscope immunolabeling are entirely relevant to electron immunocytochemistry. 

Shifting from confocal microscopy to electron microscopy requires a completely different method of sample preparation and sectioning, which translates into more time and more money. A first step in preparation is light microscopic immunocytochemistry to determine the correct antibody dilutions, the proper detergent, and the needed blocking agents. Then, as needed, move to electron microscopic immunocytochemistry with the same sample. 

Yes. A thin slice of tissue can be fixed to a microscope slide and incubated with a specific labelled probe molecule that hybridizes only to the transcript or protein of interest. The probe can be complementary DNA or antisense RNA when RNA is the target, this is called in situ hybridization (ISH). Proteins in a tissue slice can be expressed as recombinant tagged fusion proteins or detected by antibodies the latter is called in situ immunocytochemistry. The label can be radioactive, such as or commonly the more precise H, fluorescent (FISH), or an enzyme that produces a colored product. The signal is then recorded by light microscopy to permit localization of the transcript or protein. 

Correlative microscopy involves multiple separate microscopy platforms, which are correlated to view the same exact structure of interest with complimentary information [10]. Such systems allow structure-function relationships to be determined at high resolutions. Examples of correlative microscopy include light and electron microscopy, light and atomic force microscopy, immunocytochemistry, and histochemistry, etc. 

Key words Immunocytochemistry, Nervous system. Light microscopy. Electron microscopy. 

EM electron microscopy, FRP fluorescent reporter protein, ICC immunocytochemistry, LM light microscopy, UV ultraviolet... 

Bonnard, C., Papermasteg D.S., and Kraehenbuhl, J.-P. (1984) The streptavidin-biodn bridge technique Application in light and electron microscope immunocytochemistry. In Immunolabelling for Electron Microscopy, (J.M. Polak, and I.M. Varndell, eds.), pp. 95-111. Elsevier, New York. 

Harris N. Immunocytochemistry for light and electron microscopy, in Plant Cell Biology (Harris N, Oparka KJ, eds.), IRL Press, Oxford, UK, 1994, pp. 157-176. 

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... 

C. Bonnard, D. S. Papermaster, and J.-P. Kraehenbuhl, The Streptavidin-biotin Bridge Technique Application in Light and Electron Microscope Immunocytochemistry, in Immunolabeling for Electron Microscopy (eds. J. M. Polak and I. M. Varndell), Elsevier, Amsterdam, 1984, pp. 95-111. 

This book, from its initial conception, had obviously to be limited in the choice of subjects, but we believe it represents a valuable and readily reproducible collection of established and emerging techniques. Such a collection is preceded by a general introductory chapter (Chap. 1) that recalls the history of immunocytochemistry, its basic principles, and its application to the study of neuronal complexity. The methods presented include immunocytochemical localization at light and electronic levels, biochemical characterization, and functional analysis in vivo or ex vivo by novel types of microscopy, as well as protocols for development and production of genetic probes. Although this book is primarily devoted to approaches for analysis of the mammalian brain, a few nonmammalian species are also taken into consideration to demonstrate the importance of alternative animal models in a more comprehensive analysis of central and peripheral neurons. 

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