螢光能量共振轉移分析
Pixel-precise bleaching & impressive heat maps


藉由螢光蛋白CFP及YFP兩分子間產生的能量轉移現象,分析蛋白質結構上的變化。首先經由顯微鏡所拍攝出樣本的影像後,使用ZEISS ZEN影像軟體分析所拍攝之影像,再從數據結果中判斷蛋白質是否有產生結構上的改變,此軟體提供3種不同的運算方式 (如:Gordon、Youvan及Xia),可精準地分析出相關數據,幫助判斷蛋白質之間交互作用關係。
更詳細的螢光共定位軟體操作分析影片(Intermolecular FRET Walkthrough, Youvan Method):
https://www.zeiss.com/microscopy/us/local/zen-knowledge-base/zen.html
文章來源: microscopy@zeiss.com

Upgrade your ZEISS microscope
Upgrade your ZEISS microscope with the FRET option and benefit from pixel-precise bleaching and impressive heat maps.
As technical requirements may apply on some systems, please contact us to learn more about FRET upgrade and how your process will benefit from an upgrade:
microscopy@zeiss.com
1.jpg
Sample image of Hep G2 cells, LSM 800 application image for FRET, two interacting proteins (donor false-colored in green, acceptor false-colored in red) in HepG2 cells ("before bleach").

Förster Resonance Energy Transfer (FRET) for ZEISS ZEN Imaging Software is an improved module to observe interactions of molecules.
With FRET two differently strained molecules will transfer energy at a distance of less than 10nm, such as that the second fluorophore, which is not excited with light, will start emitting light. Through heat maps based on the most commonly used models: Gordon, Youvan, and Xia, FRET enables to demonstrate its efficiencies at structural changes of molecules (protein folding).

Highlights
Pixel-precise bleaching
Define your ROIs with pixel-precise for Acceptor Photobleaching (AP)

Two steady-state techniques
Benefit from two steady-state techniques at disposal: Acceptor Photo-bleaching (AP) and Sensitized Emission (SE)

Heat maps
Display your FRET efficiencies by heat maps based on the three most commonly used models

FRET for ZEISS ZEN Imaging Software
Impressive heat map

 
2.jpg
Sample image of two interacting proteins tagged with CFP (donor) and YFP
(acceptor) in HepG2 cells. Left panel shows the donor. Right panel indicates
the FRET channel obtained by sensitized emission from which the FRET
efficiencies calculated according to Youvan are displayed as fraction.






 

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