Measurement of Three-Dimensional Anisotropic Diffusion by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0063-WXH)

Introduction

Fluorescence recovery after photobleaching (FRAP) is a versatile and widely used tool for the determination of local diffusion properties within solutions, cells, tissues, and biomaterials. Due to the high spatial resolution, FRAP offers the possibility to microscopically examine a specific region of a sample.
More importantly, a variety of FRAP techniques has been developed to understand the relationship between tissue structure and solute diffusion properties by determining the diffusional anisotropy in the biological tissues. For example, the fluorescence imaging of continuous point photobleaching (FICOPP), elliptical surface photobleaching, line FRAP and spatial Fourier analysis (SFA) based FRAP techniques have been utilized to investigate the diffusional anisotropy of different solutes in varying types of tissues (e.g., the cartilaginous tissues and fibrosarcoma tumors). It was found that the structure and orientation of collagen fibers in the tissue extracellular matrix (ECM) leads to diffusion anisotropy.

Add to Cart Please Inquire

Principle Applications Procedure Materials Notes Related Products

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.

Other recommended products

Study of Transcription Factor Interaction with Chromatin by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0011-WXH)
Study of the biochemistry of nucleocytoplasmic exchange by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0050-WXH)
Study of SOX9 transcription factor activity by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0068-WXH)
Detection of the Diffusion in Model Membranes by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0053-WXH)
Measurement of Site-Specific Molecular Diffusion in Articular Cartilage by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0008-WXH)
Study of Cellular Signaling and Protein-Protein Interactions by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0052-WXH)
Measurement of the translation mobility of concanavalin A by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0025-WXH)
Characterization of diffusion of macromolecules in konjac glucomannan solutions and gels by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0026-WXH)