1. Provider: OLYMPUS
2. Model: FV1000MPE
3.Charge: 150 Yuan/h
1. IR pulsed laser with negative chirp for multiphoto excitation:
a.Mode-locked Ti: sapphire laser [femtosecond laser (equipped with a group velocity dispersion correction/control device)]
b.MaiTai HP DeepSee-OL: 690-1040nm
2. Visible light laser AOFT laser combiner
a. LD laser: 405nm
b. LD (R) laser: 635nm
c. Multi Ar laser: 458nm, 488nm, 514nm
d. HeNe(G) laser: 543 nm
e.Modulation: Continuously adjustable via an AOTF (0.1% to 100% in 0.1% increments)
1. Confocal detector
a. Detector: 3 channels for fluorescence detection (photomultipliers);
b. Wavelength resolution: 2nm; Wavelength switching speed: 100nm/ms;
c. Field Number:18.
2. Scanning modes
a. Pixel size: 64×64—4096×4096 pixels;
b. Scanning speed: (pixel time): 2μs-200μs;
c. High-speed scanning mode:16 frames/s (256×256);
d. Scanning Dimensions: Time, Z-axis, wavelength or their random combination;
e. Scanning in line: straight line or random line.
A motorize focus module inside the microscope is used. Minimum increment: 10 nm
Objects: 5×, 10×, 20×, 40×；60×, 100×(Oil)；10×, 20×, 25×, 40×, 60× (Water)
FV10-ASW Ver. 2.0
Main features and Service
Two-Photon Fluorescence Microscopy is a powerful research tool that combines the advanced optical techniques of laser scanning confocal microscopy with the long wavelengths of two-photon excitation. It is based on the phenomenon that at high photon density, two photons can be simultaneously absorbed (combining their energies) to provoke the electronic transition of a fluorophore to the excited state. Because the energy of a photon is inversely proportional to its wavelength, the two photons have approximately twice the wavelengths of light required for single-photon excitation. Two-photon excitation high photon density is necessary to ensure a sufficient level of fluorophore excitation. To prevent photo-damage, the two-photon microscope utilizes high-power mode-locked pulsed laser, which generates a significant amount of power during pulse peaks, but has an average power that is sufficiently low as not to damage the specimen. A typical pulsed laser configuration employs short duty cycles of around 100 femtoseconds with a repetition rate of 80 to 100 megahertz. Because of high numerical aperture objectives, two-photon excitation occurs only at the focal point of a diffraction-limited microscope, a detecting pinhole is not need in two-photon microscope, thereby increasing detection efficiency. Two-photon microscopy has many advantages over confocal microscopy:
1.Long wavelength infrared light scatters less than light of shorter wavelengths and penetrates through specimen more easily;
2.lack of absorption from fluorophores positioned outside the focal plane allows more excitation light to penetrate through the specimen and reach the plane of focus;
3. long wavelength infrared laser is less phototoxic than lasers of shorter wavelengths;
4. photobleaching and phototoxicity occurs only at the focal point.
For these reasons, two-photon microscopy is particularly suitable for experiments including long term live cell imaging, imaging in deep tissues and photobleaching or other manipulations in small localized regions.