Microscopy techniques such as FRAP, FRET and FLIP are increasingly being used to investigate intracellular protein movement. With Fluorescece recovery after photobleaching the target area is bleached for a fixed amount of time and the recovery of fluorescence in that area is used to determine protein movement, motility and dynamics. In Fluorescence loss in photobleaching bleaching is applied throughout the experiment and the movement of molecules is assessed by measuring the fluorescent change in the area surrounding the target region. This loss of fluorescence is due to molecules moving into the bleached area.

           

The FV 1000_1 has two laser scanners, one for confocal imaging and the other for simultaneous stimulation ( 405 laser). They can be illuminated separately and independently, making it possible to stimulate the specimen during observation. As a result, the rapid cell reactions that occur right after laser stimulation can be accurately and reliably captured.(The above image shows the blue 488nm image scanning laser and the purple 405nm bleaching laser.1 frames 512x512 normal scanning and then bleaching with the 405nm laser.) Our Nikon A1R and our FV1000_ is also capable of simultaneous scanning and FRAP. On these systems  any laser line can be used for your photo-activation and FRAP experiments.

 

 

 
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Above :Demonstration of FRAP and of SIM scanning. Image 2  demonstrates the SIM scanner on FV1000  and was taken by removing the objective lens and directing lasers onto the ceiling.

 

 Above Image demonstration FRAP and show recovery time. The images were taken using the Olympus the FV1000.( I. Konig)

Useful links:

Introduction to FRAP by K. Miura Molecular Dynamics 2004

Overview of FRAP www.embl-heidelberg.de

Precise calculation of Photoactivation kenetics( Nature Methodes dec 2006)

Higher optical Fluorescent protiens Olympus Java tutorial

 

 

 


 

 

 

 


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Last modified: 02/25/16