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TIRF Microscope System

Photonics.com
Oct 2007
Leica Microsystems GmbHRequest Info
 
For Live-Cell Research
WETZLAR, Germany, Oct. 25, 2007 -- The new Leica TIRF (total internal reflection fluorescence) microscope system, called the Leica AM TIRF MC, allows the entire fluorescence excitation spectrum to now be used for real-time visualization of live cell dynamics, according to manufacturer Leica Microsystems GmbH.

Leica's scanner sensor automatically matches the TIRF angle at all wavelengths to the required penetration depth, and positions the laser so that the TIRF penetration depth remains constant even when the wavelength is changed. Researchers using Leica's TIRF technology benefit from a unique level of convenience as well as from reproducible scientific results, the company said.

LeicaTIRF.jpgThe Leica AM TIRF MC integrates four solid-state lasers for the excitation of fluorophores at wavelengths from 405 to 632nm. The system features extremely short switching times and an ultrahigh synchronized frame rate. For the scientist, Leica Microsystems said, this means excellent flexibility for experiments with multiple fluorophores. The system is particularly useful for exploring molecular interaction at cell membranes and molecular interaction of proteins and receptors involved in transport mechanisms. The system lets a scientist examine single molecules near the cell membrane, study colocalization and vesicle transport, and combine TIRF and fast FRET analysis.

The unit provides wavelengths of 405, 488, 561, and 632 nm for multicolor applications. Individual laser lines are paired and controlled quickly and precisely via acousto-optical tunable filters during a switching time of only 1 ms.

Leica's newly designed SyncBoard controls the fast frame rate of up to 30 fps. This high temporal resolution for the excitation of multiple fluorophores, combined with the high spatial TIRF resolution of 70-300 nm in the z-axis, are key prerequisites for visualizing transport processes in cell membranes or protein interactions in real time.

Even individual molecule interaction, kinetics, and colocalization can be visualized and measured with Leica's highly sensitive TIRF system. Maintaining constant penetration depth when the laser lines are switched is an essential requirement for the use of multiple fluorophores. With an excellent signal-to-noise ratio, the entire dynamic range of the high-sensitivity Leica EMCCD (electron multiplying CCD) camera can be used. Leica said its camera takes top quality pictures even of weak fluorescence signals.

"The Leica AM TIRF MC offers live cell researchers leading-edge technology that reliably provides reproducible scientific results due to its automatic control of penetration depth," said Leica Marketing Manager Werner Kampe. "In addition, the system is highly versatile and extremely easy to use."

On the practical side, the Leica AM TIRF MC is a compact system with a small footprint and is a safety class 1 laser unit that uses a quiet air-cooling system, the company said. All four laser lines are guided via a single multimode fiber optic cable. The TIRF module can be connected to Leica's inverted microscopes as well as to the high-end Leica TCS SP5 confocal system.

For more information, visit: www.leica-microsystems.com

Leica Microsystems GmbH
Ernst-Leitz-Strasse 17-37
35578 Wetzlar
Phone: +49 6441-29-0
Fax: +49 6441-29-2590



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GLOSSARY
cell
1. A single unit in a device for changing radiant energy to electrical energy or for controlling current flow in a circuit. 2. A single unit in a device whose resistance varies with radiant energy. 3. A single unit of a battery, primary or secondary, for converting chemical energy into electrical energy. 4. A simple unit of storage in a computer. 5. A limited region of space. 6. Part of a lens barrel holding one or more lenses.
excitation
1. The process by which an atom acquires energy sufficient to raise it to a quantum state higher than its ground state. 2. More specifically with respect to lasers, the process by which the material in the laser cavity is stimulated by light or other means, so that atoms are converted to a semistable state, initiating the lasing process.
fluorescence
The emission of light or other electromagnetic radiation of longer wavelengths by a substance as a result of the absorption of some other radiation of shorter wavelengths, provided the emission continues only as long as the stimulus producing it is maintained. In other words, fluorescence is the luminescence that persists for less than about 10-8 s after excitation.
microscope
An instrument consisting essentially of a tube 160 mm long, with an objective lens at the distant end and an eyepiece at the near end. The objective forms a real aerial image of the object in the focal plane of the eyepiece where it is observed by the eye. The overall magnifying power is equal to the linear magnification of the objective multiplied by the magnifying power of the eyepiece. The eyepiece can be replaced by a film to photograph the primary image, or a positive or negative relay...
photonics
The technology of generating and harnessing light and other forms of radiant energy whose quantum unit is the photon. The science includes light emission, transmission, deflection, amplification and detection by optical components and instruments, lasers and other light sources, fiber optics, electro-optical instrumentation, related hardware and electronics, and sophisticated systems. The range of applications of photonics extends from energy generation to detection to communications and...
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