Controlled Light Exposure Microscopy (CLEM) : New Imaging Approach Reduces Photobleaching and Phototoxicity

✍ Scribed by R. A. Hoebe; C. H. Van Oven; T. W. J. Gadella Jr; P. B. Dhonukshe; C. J. F. Van Noorden; E. M. M. Manders

• Book ID: 102866574
• Publisher: Wiley (John Wiley & Sons)
• Year: 2007
• Weight: 841 KB
• Volume: 9
• Category: Article
• ISSN: 1439-4243
• DOI: 10.1002/imic.200790117

✦ Synopsis

Photobleaching and phototoxicity are often the Achilles' heels of fluorescence live-cell imaging. Controlled Light Exposure Microscopy (CLEM) is a novel, simple imaging approach that reduces photobleaching and phototoxicity 2 to 10-fold without compromising image quality. The basic concept of CLEM is that light is only used where it is needed by spatial control (pixel by pixel) of the light exposure time. Experiments show that CLEM results in a 2 to 10-fold reduction of photobleaching and phototoxicity. As a bonus, CLEM increases the dynamic range of the fluorescence intensity at least by a factor of two.

ple is a matter of finding the balance be tween S/N and phototoxicity and photo bleaching (fig. ). One thing is clear: you can't have it all! CLEM is a new technology that breaks this balance. Controlled Light Exposure Microscopy (CLEM) enhances cell viabil ity without deteriorating image quality. The basic concept of CLEM is that excita tion light is used only where it is needed by spatial control (pixel by pixel) of the light exposure time [1-3].