Olympus has introduced the Fluoview FV1000-MPE multiphoton laser scanning microscope system for deep observation and imaging of living or thickly sliced specimens
Neuroscientists and cell biologists can use the Fluoview FV1000-MPE multiphoton laser scanning microscope system to image dynamic processes up to several hundred microns deep in living cells and tissues without inflicting significant photo-damage to specimens.
The system is particularly useful for repeated, long-range or time-lapse exposures, and any application that requires deep imaging and/or extensive fluorescence excitation.
The FV1000-MPE incorporates the use of a femtosecond pulsed laser to achieve photon density at the focal plane with minimal damage or photobleaching of living cells.
It is optimised for use with near-infrared (IR) wavelengths, and a full line of water immersion objectives is available for the best possible specimen penetration and imaging.
In multiphoton microscopy, a fluorescently-tagged specimen absorbs the energy from two (or more) photons that arrive simultaneously, and emits a fluorescence signal as if having been stimulated by a photon of double (or triple) the photon energy.
Multiphoton systems can use excitation wavelengths that are of a longer, lower energy type, which scatter less as they pass through tissue.
In addition, the detector is non-descanned (ie, light does not need to travel through the scanning mirrors or confocal aperture before reaching the detector) and can thus be located as close to the objective as possible for maximum efficiency.
Multiphoton systems allow researchers to achieve thinner optical sectioning and also cause less photobleaching and phototoxicity to surrounding areas of the specimen.
They are thus ideal for deep, in vivo imaging.
The system is built around Olympus's widely used FV1000 confocal imaging platform, with its proprietary SIM scanner.
This allows for two lasers to be used simultaneously in different regions of the specimen.
When combined with the MPE system, one can use two multiphoton lasers simultaneously for 3D uncaging experiments.
There are two specialised optics sets available in addition to the standard in vivo observation optics set.
The acousto-optic module (AOM) set allows high-speed on-off toggling of the laser.
Negative chirp, which corrects the time delay of laser pulse components and adjusts the beam so that the narrowest pulse is achieved, is provided as standard with the AOM optics.
Additionally, there is a SIM scanner optics set, which delivers highly localised stimulation of a specific site in three dimensions, making 3D uncaging possible.
The optics integrated with this set are also equipped with an AOM module, for highly precise two-dimensional stimulation of the targeted area.
