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. 2022:2422:131-146.
doi: 10.1007/978-1-0716-1948-3_9.

Fluorescent Immunohistochemistry

Affiliations

Fluorescent Immunohistochemistry

Vanessa Moreno et al. Methods Mol Biol. 2022.

Abstract

Immunofluorescence is an important immunochemical technique that utilizes fluorescence-labeled antibodies to detect specific target antigens. It is used widely in both scientific research and clinical laboratories. Immunofluorescence allows for excellent sensitivity and amplification of signal in comparison to immunohistochemistry. However, analysis of samples labeled with fluorescence-labeled antibodies has to be performed using a fluorescence microscope or other type of fluorescence imaging. There are two methods available: direct (primary) and indirect (secondary) immunofluorescence. Here, we describe the principle of immunofluorescence methods as well as the preparation of fresh-frozen and formalin-fixed, paraffin embedded tissues for both direct and indirect immunofluorescence labeling.

Keywords: Direct immunofluorescence; FFPE; Fresh-frozen; Immunofluorescence; Immunohistochemistry; Indirect immunofluorescence.

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References

    1. Im K, Mareninov S, Diaz MFP, Yong WH (2019) An introduction to performing immunofluorescence staining. Methods Mol Biol 1897:299–311 - PubMed - PMC
    1. Coons AH, Creech HJ, Jones RN, Berliner E (1942) The demonstration of pneumococcal antigen in tissues by the use of fluorescent antibody. J Immunol 45:159–170
    1. Coons AH (1961) The beginnings of immunofluorescence. J Immunol 87:499–503 - PubMed
    1. Coons AH, Kaplan MH (1950) Localization of antigen in tissue cells; improvements in a method for the detection of antigen by means of fluorescent antibody. J Exp Med 91:1–13 - PubMed - PMC
    1. Burkholder PM (1961) Complement fixation in diseased tissues : I. fixation of Guinea pig complement in sections of kidney from humans with membranous glomerulonephritis and rats injected with anti-rat kidney serum. J Exp Med 114:605–616 - PubMed - PMC

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