Analysis of chromatin structure in preimplantation embryos by Fluorescence recovery after photobleaching (FRAP) (CAT#: STEM-MT-0033-WXH)

Name: Analysis of chromatin structure in preimplantation embryos by Fluorescence recovery after photobleaching (FRAP)
SKU: STEM-MT-0033-WXH
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Introduction

Embryos at the 1-cell stage are totipotent and this totipotency is gradually lost during preimplantation development. It was suggested that chromatin structure is tightly associated with the differentiation state of cells. Numerous reports have shown that the chromatin structure of pluripotent embryonic stem (ES) cells is in a “loose” or “open” state, and changes to “tight” or “compact” when they differentiate.

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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.