TIRF Microscope Illuminator Offers Simultaneous 4-Channel Image Capture

A total internal reflection fluorescence (TIRF) microscopy illuminator offers four motorized channels for simultaneous image capture and intuitive, user-friendly software control of TIRF parameters. The system also allows easy transition back and forth to widefield fluorescence and space-saving ergonomic design.

TIRF imaging was developed to deliver information about the surfaces of cells where vital functions such as communication, absorption, signaling, growth, and movement occur. TIRF imaging occurs where the cell membrane comes in contact with a cover slip. The laser illumination light's angle of incidence dictates how deeply an evanescent wave penetrates into the cell, and lasers of different wavelengths require varying angles to operate consistently with each other.

The new cell^TIRF system has four individually controlled motorized laser inputs for TIRF imaging. Each laser wavelength is optimally focused and each angle is individually set, allowing different wavelengths to have the same penetration depth. Thus the cell^TIRF system can simultaneously capture multiple channels with independently adjusted TIRF angles.

The software is operated through a simple graphical user interface (GUI), keyboard arrow keys or the mouse wheel, making it easy to control the incident angle of each wavelength and adjust TIRF penetration depths. Calculated penetration depths can be preset for all four lasers with a single mouse click; the system will individually adjust each laser's angle automatically to simultaneously capture TIRF from all four channels.

The system has all four laser inputs coming in from one side, making it easy to set up and integrate with incubator systems. Clean-up filters for lasers can be inserted quickly if needed. In addition, the main unit is built from single billet aluminum for rigidity and robustness, making it more tolerant of small temperature fluctuations that may occur in rooms where it is placed.

One laser line can be adjusted so that the system can do fluorescence recovery after photobleaching (FRAP) experiments. A handy button is available to switch to widefield imaging, allowing users to visualize the complete cell profile, see nuclei, or find the field they want to observe. TIRF imaging mode can be reestablished in less than 500 msec. Users can also control laser intensities easily onscreen.

Olympus (center Valley, PA, USA) launched the cell^TIRF microscopy illuminator together with new Olympus laser systems incorporating the most commonly used laser wavelengths in the 405 nm to 640 nm range. The directly coupled laser systems feature attenuable laser power up to 100 mW, and offer better delivery efficiency to the microscope than laser combiner options. Each laser system is a small, stackable integrated unit.

"Olympus was the first company to commercialize TIRF microscopy, and as the pioneer in this field, we've taken another key step with advanced motorization, additional inputs for simultaneous TIRF acquisition, and a great GUI to easily and repeatably set incident angles,” said Stuart Shand, TIRF marketing manager for Olympus America Inc.

Olympus America Scientific Equipment Group provides microscope imaging solutions for clinicians, doctors, researchers, and educators. The company's microscope systems offer excellent optics, superior construction and system versatility to meet the ever-changing needs of microscopists, paving the way for future advances in life science.

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