Let There Be Light!
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 Förster resonance energy transfer (FRET) and in situ...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2016-11-01
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| Series: | Proteomes |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2227-7382/4/4/36 |
| _version_ | 1811308175120400384 |
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| author | Doroteya Raykova Björn Koos Anna Asplund Márton Gelléri Ylva Ivarsson U. Helena Danielson Ola Söderberg |
| author_facet | Doroteya Raykova Björn Koos Anna Asplund Márton Gelléri Ylva Ivarsson U. Helena Danielson Ola Söderberg |
| author_sort | Doroteya Raykova |
| collection | DOAJ |
| description | 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 Förster 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. |
| first_indexed | 2024-04-13T09:18:33Z |
| format | Article |
| id | doaj.art-1dd337435fc94abc89d7eaed55b5cc76 |
| institution | Directory Open Access Journal |
| issn | 2227-7382 |
| language | English |
| last_indexed | 2024-04-13T09:18:33Z |
| publishDate | 2016-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Proteomes |
| spelling | doaj.art-1dd337435fc94abc89d7eaed55b5cc762022-12-22T02:52:41ZengMDPI AGProteomes2227-73822016-11-01443610.3390/proteomes4040036proteomes4040036Let There Be Light!Doroteya Raykova0Björn Koos1Anna Asplund2Márton Gelléri3Ylva Ivarsson4U. Helena Danielson5Ola Söderberg6Department of Pharmaceutical Biosciences, Pharmaceutical Cell Biology, Biomedical Center, Box 594, Uppsala University, SE-751 08 Uppsala, SwedenDepartment of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Straße 11, 44227 Dortmund, GermanyDepartment of Immunology, Genetics and Pathology, Dag Hamarskjölds väg 14B, Rudbeck Laboratory, Uppsala University, 751 85 Uppsala, SwedenDepartment of Systemic Cell Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Straße 11, 44227 Dortmund, GermanyDepartment of Chemistry-BMC, Box 576, Uppsala University, SE-751 23 Uppsala, SwedenDepartment of Chemistry-BMC, Box 576, Uppsala University, SE-751 23 Uppsala, SwedenDepartment of Pharmaceutical Biosciences, Pharmaceutical Cell Biology, Biomedical Center, Box 594, Uppsala University, SE-751 08 Uppsala, SwedenThe 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 Förster 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.http://www.mdpi.com/2227-7382/4/4/36high resolution microscopyprotein–protein interactionspost-translational modificationsFRETin situ PLAproxHCR |
| spellingShingle | Doroteya Raykova Björn Koos Anna Asplund Márton Gelléri Ylva Ivarsson U. Helena Danielson Ola Söderberg Let There Be Light! Proteomes high resolution microscopy protein–protein interactions post-translational modifications FRET in situ PLA proxHCR |
| title | Let There Be Light! |
| title_full | Let There Be Light! |
| title_fullStr | Let There Be Light! |
| title_full_unstemmed | Let There Be Light! |
| title_short | Let There Be Light! |
| title_sort | let there be light |
| topic | high resolution microscopy protein–protein interactions post-translational modifications FRET in situ PLA proxHCR |
| url | http://www.mdpi.com/2227-7382/4/4/36 |
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