Evaluation of Lateral Diffusion of Lipids in Continuous Membranes between Freestanding and Supported Areas by Fluorescence Recovery after Photobleaching
Autor: | Koji Sumitomo, Hiroshi Nakashima, Azusa Oshima |
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Rok vydání: | 2019 |
Předmět: |
Chemistry
Fluorescence recovery after photobleaching 02 engineering and technology Surfaces and Interfaces 010402 general chemistry 021001 nanoscience & nanotechnology Condensed Matter Physics 01 natural sciences 0104 chemical sciences Membrane Lateral diffusion Electrochemistry Biophysics lipids (amino acids peptides and proteins) General Materials Science 0210 nano-technology Spectroscopy Planar bilayer |
Zdroj: | Langmuir. 35:11725-11734 |
ISSN: | 1520-5827 0743-7463 |
Popis: | The lateral diffusion of lipids is a key factor when functionalizing artificial planar bilayer lipid membranes (BLMs). Fluorescence recovery after photobleaching (FRAP) is an established method for evaluating the fluidity of BLMs by the quantitative determination of the diffusion coefficient. However, a BLM with a uniform diffusion coefficient is usually assumed for analysis. In addition, when the BLM to be evaluated is small, the spread of a bleached circle caused by lateral diffusion during the bleaching process and the divergence of the laser used for bleaching interfere with the quantitative analysis. In this study, a numerical calculation was adopted to make it possible to analyze the continuous BLM between freestanding and supported areas, where the diffusion coefficients change depending on the presence or absence of an interaction with the substrate. A quantitative evaluation independent of such bleaching conditions as the bleaching diameter was also ensured by incorporating the spreading effect of the bleached circle in the calculation, which was employed to analyze a freestanding BLM with a diameter of only a few micrometers. By comparing calculations and experiments on FRAP recovery curves, we found that there are multiple diffusion elements and high diffusion barriers at the boundary between the freestanding and supported areas in a BLM over a SiO |
Databáze: | OpenAIRE |
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