Fluorescent markers emit light in a different wavelength after being excited by specific wavelengths of light. If a very intense light is used to excite the fluorophores, or they are exposed for too long, the fluorescent molecules will lose their ability to emit light. This phenomenon is called bleaching or photobleaching, and the process is an irreversible photochemical alteration. Some techniques take advantage of bleaching, such as fluorescence recovery after photobleaching (FRAP). FRAP is a technique developed by Axelrod et al. in 1976, it is a method for studying protein mobility in living cells. In FRAP, a specific area of a cell or tissue is photobleached by intense laser light to eliminate fluorescence. This area is usually the cell membrane or the area where diffusion occurs (such as the nucleus) because FRAP requires the free movement of fluorescent molecules to function. As the bleached fluorophores move out and the healthy fluorophores from other areas move in, the fluorescence of the bleached area slowly recovers. This phenomenon is called fluorescence recovery after photobleaching.
Because of these characteristics, FRAP has been widely used to determine diffusion and interaction/binding properties in biology and materials science. In cell biology, this technology can be used to study the movement of proteins and lipids in the plasma membrane, cytoplasm, and nucleus. In fields related to pharmaceuticals, food, textiles, hygiene products and cosmetics, this technology can be used to study the influence of the diffusion of solute and solvent molecules on the properties and functions of the final product.
Figure 1. The principle of FRAP (Lorén, N.; et al. 2015)
Creative Proteomics is a professional biotechnology company that has accumulated a lot of successful experience on FRAP. There are many subtleties in accurately performing FRAP experiments, and various FRAP models are based on specific assumptions, which makes it difficult for the non-specialist to choose which model is most suitable for a specific application. Our first-class expert team can customize experimental programs according to the different needs of customers, and can provide spatial FRAP models to extract more information from the samples. The speed and sensitivity of our confocal microscopes have been upgraded to make the image acquisition of the recovery process is more efficient, which can better interpret the data. In addition, we also provide excellent inverse FRAP (iFRAP) technology to help customers better study the diffusion kinetics of molecules within an immobile intracellular structure.
Customers can choose different technology platforms according to project requirements, or contact us directly for consultation, and our expert team will provide you with customized experimental procedures.
Creative Proteomics is an international biotechnology company dedicated to research in molecular interactions and other related fields. The fluorescence recovery after photobleaching (FRAP) platform we constructed has the characteristics of high quality and efficiency, and the data obtained can be directly used for paper publication. Our one-stop service aims to save customers time and money.