PMID: 8985137Dec 1, 1996Paper

Relationship between immunostaining intensity and antigen content in sections

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
J WatanabeS Kanamura

Abstract

We studied the relationship between staining intensity of immunohistochemical reaction and antigen content in sections. Alpha-fetoprotein (AFP) and albumin in sections cut from livers of newborn, 5-, 10-, 20-, and 60-day-old rats were examined as examples. First, we compared average immunostaining intensity (sum of specific absorbance in pixel/number of pixels) measured by image processing (IP), with antigen content measured by immunochemical assay to determine whether the intensity is proportional to antigen content. The intensity of AFP was proportional to the antigen content, whereas that of albumin was not. Subsequently, the antigen preservation test was carried out to determine whether the intensity was decreased by fixation and, if so, which type of decrease (proportional or disproportionate) occurred. Thereafter, antigen content in the same portion in the same immunostained section was measured by the microphotometric (MP) method followed by the IP method, because the MP method gives a low average antigen content when a decrease in antibody binding occurs in sections, whereas the average antigen content measured by the IP method is unchanged. The intensity of AFP decreased primarily by a proportional decrease in antigeni...Continue Reading

Citations

Oct 30, 2015·Journal of Anatomy·Samantha L EatonThomas M Wishart
Sep 19, 1998·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·K MinoS Kanamura
Feb 12, 1998·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·K A RothR G Lorenz
May 20, 1999·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·H SarteletD Gaillard
Jun 20, 1998·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Y AsakaS Kanamura
Jan 28, 2003·Journal of Neuroscience Research·Tomoki NakamizoShun Shimohama
Dec 12, 2018·ILAR Journal·Famke AeffnerKristin Wilson

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