Unlock Exclusive Discounts & Flash Sales! Click Here to Join the Deals on Every Wednesday!

Detection of the Diffusion in Model Membranes by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0053-WXH)

Introduction

The descriptions of the fluid-mosaic model led to an increased focus on understanding the lipid dynamics in biological membranes and gave rise to multiple ways of investigating these; a very popular technique is the use of synthetic membranes. Several synthetic membrane systems have been developed to understand the basic lipid physics in biological membranes. When working with synthetic membranes, the composition and behavior are better controlled, allowing for a more consistent investigation of the feasibility of analytical methods.




Principle

Fluorescence recovery after photobleaching (FRAP) is a microscopy technique capable of quantifying the mobility of molecules within cells. By exploiting the phenomenon of photobleaching, fluorescent mole- cules within a region of interest can be selectively and irreversibly 'turned off'. It is capable of quantifying the two-dimensional lateral diffusion of a molecularly thin film containing fluorescently labeled probes, or to examine single cells.

Applications

• Characterization of the mobility of individual lipid molecules within a cell membrane.
• Analysis of molecule diffusion within the cell
• Study of protein interaction partners, organelle continuity and protein trafficking.

Procedure

1. An initial fluorescence of fluorescent molecules is measured in the region of interest (ROI).
2. The fluorescent molecules are rapidly photobleached by focusing the high-intensity laser beam onto the defined area.
3. The exchange of bleached molecules with unbleached molecules from the surrounding region is followed over time using a low-intensity laser.

Materials

• Optical microscope.
• Light source.
• Fluorescent probe.
Advertisement