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Let There Be Light!
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.ORCID iD: 0000-0001-6452-2199
Max Planck Inst Mol Physiol, Dept Syst Cell Biol, Otto Hahn Str 11, D-44227 Dortmund, Germany..
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
Max Planck Inst Mol Physiol, Dept Syst Cell Biol, Otto Hahn Str 11, D-44227 Dortmund, Germany.;Tech Univ Dortmund, Fac Chem & Chem Biol, Otto Hahn Str 6, D-44221 Dortmund, Germany..
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2016 (English)In: Proteomes, ISSN 2227-7382, Vol. 4, no 4, 36Article, review/survey (Refereed) Published
Abstract [en]

The invention of the microscope has been fundamental for the understanding of tissue architecture and subcellular structures. With the advancement of higher magnification microscopes came the development of various molecular biology tools such as Forster resonance energy transfer (FRET) and in situ proximity ligation assay (in situ PLA) to monitor protein interactions. Microscopy has become a commonly used method for the investigation of molecular events within the cell, for the identification of key players in signaling networks, and the activation of these pathways. Multiple approaches are available for functional analyses in single cells. They provide information not only on the localization of proteins at a given time point, but also on their expression levels and activity states, allowing us to pinpoint hallmarks of different cellular identities within tissues in health and disease. Clever solutions to increase the sensitivity of molecular tools, the possibilities for multiplexing, as well as image resolution have recently been introduced; however, these methods have their pros and cons. Therefore, one needs to carefully consider the biological question of interest along with the nature of the sample before choosing the most suitable method or combination of methods. Herein, we review a few of the most exciting microscopy-based molecular techniques for proteomic analysis and cover the benefits as well as the disadvantages of their use.

Place, publisher, year, edition, pages
2016. Vol. 4, no 4, 36
Keyword [en]
high resolution microscopy, protein-protein interactions, post-translational modifications, FRET, in situ PLA, proxHCR
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-316327DOI: 10.3390/proteomes4040036ISI: 000392385500002OAI: oai:DiVA.org:uu-316327DiVA: diva2:1095992
Funder
EU, FP7, Seventh Framework Programme, 278568Swedish Research Council, 2014-02968, D0571301 , 2012-5092
Available from: 2017-05-16 Created: 2017-05-16 Last updated: 2017-05-16Bibliographically approved

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Raykova, DoroteyaAsplund, AnnaIvarsson, YlvaDanielson, U. HelenaSöderberg, Ola
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