The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane
The plasma membrane of mammalian cells is involved in a wide variety of cellular processes, including, but not limited to, endocytosis and exocytosis, adhesion and migration, and signaling. The regulation of these processes requires the plasma membrane to be highly organized and dynamic. Much of the...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-05-01
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| Series: | Membranes |
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| Online Access: | https://www.mdpi.com/2077-0375/13/5/492 |
| _version_ | 1797599147913641984 |
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| author | Charles A. Day Minchul Kang |
| author_facet | Charles A. Day Minchul Kang |
| author_sort | Charles A. Day |
| collection | DOAJ |
| description | The plasma membrane of mammalian cells is involved in a wide variety of cellular processes, including, but not limited to, endocytosis and exocytosis, adhesion and migration, and signaling. The regulation of these processes requires the plasma membrane to be highly organized and dynamic. Much of the plasma membrane organization exists at temporal and spatial scales that cannot be directly observed with fluorescence microscopy. Therefore, approaches that report on the membrane’s physical parameters must often be utilized to infer membrane organization. As discussed here, diffusion measurements are one such approach that has allowed researchers to understand the subresolution organization of the plasma membrane. Fluorescence recovery after photobleaching (or FRAP) is the most widely accessible method for measuring diffusion in a living cell and has proven to be a powerful tool in cell biology research. Here, we discuss the theoretical underpinnings that allow diffusion measurements to be used in elucidating the organization of the plasma membrane. We also discuss the basic FRAP methodology and the mathematical approaches for deriving quantitative measurements from FRAP recovery curves. FRAP is one of many methods used to measure diffusion in live cell membranes; thus, we compare FRAP with two other popular methods: fluorescence correlation microscopy and single-particle tracking. Lastly, we discuss various plasma membrane organization models developed and tested using diffusion measurements. |
| first_indexed | 2024-03-11T03:30:37Z |
| format | Article |
| id | doaj.art-afa069a58f244a758f2de7b4684bf6f5 |
| institution | Directory Open Access Journal |
| issn | 2077-0375 |
| language | English |
| last_indexed | 2024-03-11T03:30:37Z |
| publishDate | 2023-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Membranes |
| spelling | doaj.art-afa069a58f244a758f2de7b4684bf6f52023-11-18T02:24:11ZengMDPI AGMembranes2077-03752023-05-0113549210.3390/membranes13050492The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma MembraneCharles A. Day0Minchul Kang1Hormel Institute, University of Minnesota, Austin, MN 55912, USADepartment of Mathematics, Texas A&M—Commerce, Commerce, TX 75428, USAThe plasma membrane of mammalian cells is involved in a wide variety of cellular processes, including, but not limited to, endocytosis and exocytosis, adhesion and migration, and signaling. The regulation of these processes requires the plasma membrane to be highly organized and dynamic. Much of the plasma membrane organization exists at temporal and spatial scales that cannot be directly observed with fluorescence microscopy. Therefore, approaches that report on the membrane’s physical parameters must often be utilized to infer membrane organization. As discussed here, diffusion measurements are one such approach that has allowed researchers to understand the subresolution organization of the plasma membrane. Fluorescence recovery after photobleaching (or FRAP) is the most widely accessible method for measuring diffusion in a living cell and has proven to be a powerful tool in cell biology research. Here, we discuss the theoretical underpinnings that allow diffusion measurements to be used in elucidating the organization of the plasma membrane. We also discuss the basic FRAP methodology and the mathematical approaches for deriving quantitative measurements from FRAP recovery curves. FRAP is one of many methods used to measure diffusion in live cell membranes; thus, we compare FRAP with two other popular methods: fluorescence correlation microscopy and single-particle tracking. Lastly, we discuss various plasma membrane organization models developed and tested using diffusion measurements.https://www.mdpi.com/2077-0375/13/5/492fluorescence recovery after photobleachingFRAPdiffusionplasma membranemicrodomainactin |
| spellingShingle | Charles A. Day Minchul Kang The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane Membranes fluorescence recovery after photobleaching FRAP diffusion plasma membrane microdomain actin |
| title | The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane |
| title_full | The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane |
| title_fullStr | The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane |
| title_full_unstemmed | The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane |
| title_short | The Utility of Fluorescence Recovery after Photobleaching (FRAP) to Study the Plasma Membrane |
| title_sort | utility of fluorescence recovery after photobleaching frap to study the plasma membrane |
| topic | fluorescence recovery after photobleaching FRAP diffusion plasma membrane microdomain actin |
| url | https://www.mdpi.com/2077-0375/13/5/492 |
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