Immunogold quantification of amino acids and proteins in complex subcellular compartments

Nature Protocols
L H BergersenVidar Gundersen

Abstract

An increasing number of imaging techniques are in use to study the localization of molecules involved in cell-to-cell signaling. Here we describe the use of immunogold procedures to detect and quantify molecules on electron micrographs. To measure the areas of the subcellular compartments under investigation, the protocol uses an overlay screen with an array of regularly spaced points. On the basis of this, the densities of the gold-labeled molecules can be calculated. Despite the limited lateral resolution of the immunogold method as used by many investigators ( approximately 30 nm), it is possible to measure the content of molecules associated with tiny tissue compartments, e.g., synaptic vesicles and different types of membrane, such as plasma membranes and vesicle membranes. The quantification protocol can be carried out without using computer programs. The entire protocol can be completed in approximately 15 d.

References

Jan 1, 1991·Progress in Neurobiology·J P Royet
Jun 15, 1991·Journal of the Autonomic Nervous System·V A Maisky, N Z Doroshenko
Dec 1, 1990·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·J Storm-Mathisen, O P Ottersen
May 1, 1988·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·H J GundersenA Vesterby
May 17, 2001·Proceedings of the National Academy of Sciences of the United States of America·A DidierP M Lledo
Aug 20, 2003·The European Journal of Neuroscience·Linda BergersenVidar Gundersen
Dec 24, 2005·Nature·Ragnhildur KáradóttirDavid Attwell
Jan 20, 2007·Nature Chemical Biology·Jyoti K Jaiswal, Sanford M Simon
Feb 21, 2007·Nature Neuroscience·Pascal JourdainAndrea Volterra
Aug 4, 2007·The European Journal of Neuroscience·I TonazziniL H Bergersen

❮ Previous
Next ❯

Citations

Jun 16, 2012·Cellular Oncology (Dordrecht)·Chang ChengLinda H Bergersen
Dec 12, 2012·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Cecilie MorlandVidar Gundersen
Jan 13, 2010·Molecular and Cellular Biology·Knut H LauritzenArne Klungland
Jun 24, 2011·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·Francis W FlynnZhaojie Zhang
May 25, 2011·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Max LarssonVidar Gundersen
Aug 20, 2011·PloS One·Runhild GammelsaeterVidar Gundersen
Jan 14, 2011·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Johanne E RinholmDavid Attwell
Jun 20, 2014·PloS One·Shubha A Dighe, Keith G Kozminski
Jul 16, 2013·Brain Structure & Function·Mats Julius StensrudVidar Gundersen
Jul 11, 2012·Basic Research in Cardiology·Arkady RutkovskiyJarle Vaage
Feb 26, 2016·Frontiers in Synaptic Neuroscience·Agnieszka Münster-WandowskiGudrun Ahnert-Hilger
Aug 5, 2009·The Journal of Physiology·Azar OmraniFiorenzo Conti
Sep 12, 2012·Neuroscience Letters·Cecilie MorlandVidar Gundersen
Aug 30, 2008·Neuropharmacology·Elena V RomanovaJonathan V Sweedler
Jun 18, 2008·Acta Neurologica Scandinavica. Supplementum·V Gundersen
Oct 6, 2011·The Anatomical Record : Advances in Integrative Anatomy and Evolutionary Biology·Hisashi FujiokaCharles L Hoppel
Oct 20, 2011·Glia·Lasse OrmelVidar Gundersen
May 27, 2015·The Journal of Comparative Neurology·Mats Julius StensrudVidar Gundersen
Sep 29, 2011·Current Opinion in Neurobiology·Kristina D Micheva, Marcel P Bruchez
Mar 22, 2014·International Journal of Radiation Biology·Nina Jeppesen EdinErik Olai Pettersen
Jan 1, 2014·Brain Structure & Function·Kaja NordengenVidar Gundersen
Dec 17, 2009·Medicine and Science in Sports and Exercise·Truls RaastadJostein Hallén
Apr 29, 2015·The Journal of Biological Chemistry·Daniel G DeMartiniDaniel E Morse
Oct 4, 2017·Scientific Reports·Nima Borhan FakouriLene Juel Rasmussen

❮ Previous
Next ❯

Related Concepts

Related Feeds

Astrocytes

Astrocytes are glial cells that support the blood-brain barrier, facilitate neurotransmission, provide nutrients to neurons, and help repair damaged nervous tissues. Here is the latest research.