CLEM | CLEM

Key Words: confocal, fluorescence imaging, lasers, Live cell imaging, photobleaching, photodamage, phototoxicity, time-lapse imaging

Definition:CLEM is a microscopy technique designed to minimize the light required for excitation in fluorescence microscopy without compromising image quality.

TECHNOLOGY:

In CLEM illumination of the fluorescent sample is determined on a per pixel basis (i.e. when and where required) by an integrated feedback process in the digital camera. Excitation light is reduced using two strategies:

The first is based on the principle that if there is no signal, then no illumination is required (for example, when imaging the background).

The second detects whether there is sufficient signal to acquire an image. If so, illumination is stopped.

APPLICATIONS:

CLEM is ideal for live imaging as it helps to reduce photobleaching and phototoxicity; the two main limitations in live-cell microscopy. Reduced fluorescence recovery times and increased cell survival offer greater flexibility in the design of time-lapse and other live cell imaging studies.

MICROSCOPE CONFIGURATION:

• CLEM is suitable for both confocal and standard epi-fluorescence techniques.
• CLEM can be configured with Nikon's C1 confocal microscopes systems, Nikon's inverted TE2000 series and Nikon's upright Eclipse i-series microscopes.
• CLEM is not suitable for non-fluorescence applications.

RECOMMENDED SYSTEM:

e.g. CLEM + Nikon C1 + Plan Fluor / Super Fluor / VC series objectives.

LINKS:

Controlled Light Exposure Microscopy Application Note