Olympus, the
world leader in objective-based total internal reflection fluorescence (TIRF)
microscopy, has announced another significant leap forward in multicolor TIRF
imaging. The Olympus cell^TIRF™ illuminator offers four motorized channels
for simultaneous image capture; intuitive, user-friendly software control of
TIRF parameters; and instant setting and confirmation of the precise TIRF angle.
The system also allows easy transition back and forth to widefield fluorescence
and a sleek, space-saving ergonomic design.
The new system has four individually controlled motorized laser inputs for
TIRF imaging. With the cell^TIRF system, each laser wavelength is optimally
focused and each angle is individually set, allowing different wavelengths to
have the same penetration depth. Combined, these features make cell^TIRF the
only such system that can simultaneously capture multiple channels with independently
adjusted TIRF angles. Researchers do not have to make time-consuming adjustments
during experiments.
Users can operate the software via a simple graphical user interface (GUI),
keyboard arrow keys or the mouse wheel, making it easier than ever to control
the incident angle of each wavelength and adjust TIRF penetration depths. Best
of all, users can preset calculated penetration depths for all four lasers with
just a single mouse click; the system will individually adjust each laser’s
angle automatically to simultaneously capture TIRF from all four channels.
A number of additional capabilities make the new system especially useful for
cell biologists and other researchers. For instance, one laser line can be adjusted
so the system can do fluorescence recovery after photobleaching (FRAP) experiments.
A handy button is available to seamlessly switch to widefield imaging, allowing
researchers 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 500msec.
Users can also control laser intensities easily onscreen.
The sleek, elegant design has all four laser inputs coming in from one side,
making the compact system easy to set up and integrate with incubator systems.
Researchers can quickly insert clean-up filters for lasers 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.
Accompanying the launch of the cell^TIRF illuminator are new Olympus laser
systems incorporating all the most commonly used laser wavelengths in the 405
– 640nm range. The directly coupled laser systems feature attenuable laser
power up to 100mW, 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.
The introduction of the cell^TIRF system builds on Olympus’ market-leading
tradition in TIRF. Olympus introduced the first TIRF microscope objective in
the U.S. in 1997, and continues to offer the largest line of TIRF optics, including
the unmatched APO 100x NA 1.65 objective that can achieve penetration depths
below 50nm. Other TIRF objectives include the APO N 60x NA 1.49, PlanApo 100x
NA 1.45, and U-APO 150x NA 1.45. The new system can be retrofitted to Olympus
IX2® microscopes including the IX71 and IX81 microscopes.
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.
Olympus’ ability to do simultaneous multichannel image capture allows
scientists to efficiently study rapidly changing intracellular processes, do
high-precision ratio experiments, and more.