Leica SP5 X Confocal/Spectral Imaging Microscopy System with White Light Laser
The white light laser-based SP5X system is a significant advance in confocal microscopy. With respect to the multiuser core imaging facility, the critical advantage of this tunable system is that the investigators no longer need to select fluorophores based on the laser lines available on the microscope. Instead, the investigators can choose the fluorophores that are optimal for the experimental approach, and tune the microscope to the experiments. The white light laser SP5X system, especially when configured with a 405 nm laser line, has the capability of covering almost all fluorophores available for biological and clinical imaging. This system uses an AOBS option and does not require any band pass filters. We use the unique capabilities of the tunable system to characterize many different potential FRET pairs that cannot be optimally used for conventional or 2photon microscopy. A key advantage of the Leica system is that we can select the optimal excitation wavelength and power to avoid photobleaching the samples. The Leica AF software has many functions, including Multicolor+3D+time, ratio imaging, FRET, FRAP, sequential scanning to avoid crosstalk, ROI real time logical operation display for calcium imaging, physiological measurements, various image analysis functions (e.g. 3D reconstruction, linear unmixing), etc.
The Leica SP5 X imaging system coupled to DMI6000 epi-fluorescence microscope equipped with various components as listed below-
- Objective lenses: 10X/0.40,
20X/0.70, 40X/1.25 (Oil, Ph), 63X/1.20 (Water), 63X/1.40-0.60 (Oil), 100X/1.40-0.70 (Oil).
- UV laser 405 nm for imaging UV fluorophores and photoactivation.
- Argon-ion laser 458 (for cyan dyes), , 488, 496, 514 nm.
- White light laser gives any wavelength from 470 nm to 670 nm in 1 nm increments.
- Six PMT detectors (5 fluorescence channels (3 conventional PMT and 2 hybrid PMTs) + 1 transmission channel).
- Conventional scanner (100 Hz - 1400 Hz).
- Temperature controlled Chamber and Eppendorf microinjection device.