Study of Self-Diffusion in Chitosan Networks by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0057-WXH)

Name: Study of Self-Diffusion in Chitosan Networks by Fluorescence recovery after photobleaching (FRAP)
SKU: STEM-MT-0057-WXH
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Introduction

Chitosan is a natural linear cationic polysaccharide obtained by N-deacetylation of chitin. The numerous physicochemical and biological properties of chitosan such as biocompatibility, non toxicity, mucoadhesion, absorption enhancement and pHdependent swelling properties are particularly interesting for many applications in the biomedical and the pharmaceutical fields. This is why chitosan networks are already used as hydrophilic matrix formers in controlled drug delivery systems.

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Principle Applications Procedure Materials

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.