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Optical system for enhanced multiphoton performance
The FV1000MPE offers aberration correction over a wide range from the visible spectrum to near-infrared, thanks to its integrated optical design featuring a dedicated IR laser port, pupil projection lens and objectives. Because every optical element was designed with multiphoton imaging in mind, the optical system functions efficiently, and allows better optical penetration when using near-infrared lasers. Olympus provides a full line of water immersion objectives designed for near-infrared imaging. |
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Efficient fluorescence acquisition via non-descanned detector system
With regular confocal systems, returning fluorescence emissions traveling from deep within a specimen can become dispersed by the specimen and degrade acquisition efficiency. The FV1000MPE provides more efficient detection with minimal loss using an external two-channel photomultiplier detection system placed close to the specimen. The system provides effective fluorescence detection without the need for a pinhole for de-scanning, because fluorescence excitation occurs only in the focal plane. Dichromatic mirrors are used in the light path to separate the light going to the two photomultiplier tubes; the mirrors and filters can be changed easily according to the specimen’s fluorescence emission characteristics. |
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Use of a femtosecond pulsed IR laser
In multiphoton excitation, a pulsed laser is used to increase photon density in the focal plane. Fluorescent molecules can be efficiently detected even at low emission levels by using a femtosecond pulsed laser. Either a Mai Tai® (Spectra-Physics, a unit of Newport Corporation) or Chameleon™ (Coherent, Inc.) laser with PC-controlled wavelength adjustment can be adapted. |
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Simultaneaous imaging and laser stimulation
The FV1000MPE can handle applications requiring multiple points or regions of interest through high-speed On/Off toggling of the laser with an acousto-optic modulator (AOM). Laser light stimulation is not limited by imaging settings as the FV1000's SIM scanner for laser light stimulation is independent of the scanner for observation. A multiphoton laser allows simultaneous manipulation of the same focal plane as the one used in imaging. |
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Optimal pulse width
Microscope optics cause delays in laser pulse components (dispersion), which in turn can create problems with multiphoton imaging. By narrowing the pulse width of the laser (by creating a negative chirp), the beam emerging from the objective is adjusted so that the optimal width of the pulse is nearly restored. This negative chirp results in ultra-short pulses of light for multiphoton excitation and helps to reduce specimen damage. |
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Optimal for in vivo observation
