Jens Randel Nyengaard

A stereologic approach to estimate the number of immunogold-labeled molecules in cells of tubules

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

OBJECTIVE: The quantity of molecules can be measured very precisely by molecular biological methods, but the capabilities of these are limited to measure only the total mass of tissue. For estimating the number of molecules at the cell level, it is necessary to combine an immunohistochemical protocol with designed-based principles of stereology at the level of electron microscopy (EM). This article focuses on the problems and practical solutions of fitting together immunohistochemistry, stereology, and electron microscopy for the estimation of the number of angiotensin II AT1 receptors in rat kidney arterioles, STUDY DESIGN: We performed the preembedding immunostaining of angiotensin II AT1 receptors using the silver-enhanced immunogold labeling system at EM level on serial sections of renal arterioles from 5 rats. RESULTS: Using this method the number of molecules can be estimated along the renal arterioles separately on the cell's surface, in cytoplasm, in nucleus, or in any subcellular location. CONCLUSION: For estimating the number of AT1 receptors, we designed a protocol that took into account the requirements for both immuno-EM and stereology. This method can be applied for estimating any molecule number in different types of cells in tubules.

Original languageEnglish
JournalAnalytical and Quantitative Cytology and Histology
Volume28
Issue1
Pages (from-to)54-60
Number of pages7
ISSN0884-6812
Publication statusPublished - 1 Feb 2006

    Research areas

  • Angiotensin II AT1 receptor, Colloidal gold, Electron microscopy, Endothelial cells, Immunohistochemistry, Kidney arterioles, Renin, Silver enhancement, Smooth muscle cells, Stereology

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