The Zeiss 980 NLO system consists of an inverted Axio Observer microscope, laser scanning confocal, 32 channels spectral imaging, one 8Y-airyscan detector and two channels non-descan (NDD) epi-fluorescence detectors, motorized piezo stage for automated scanning, definite focus for focus compensation in time-lapse imaging and an IR Insight 3X, ultrafast Ti:Sapphire laser for multiphoton excitation. The advantage of this system is the 8Y airyscan module which provides a better resolution with a faster speed (see table 2). The 32-channel descan spectral GaAsP (Gallium Arsenide Phosphide) detector with ~40% quantum efficiency with spectral resolution about 8.7 nm covers the spectral range 415-735 nm. For lambda sequential spectral scanning one can select 2.9 nm or 4.4 nm steps. There are two peltier cooled PMT detectors (located on both sides of the spectral detector) to cover the spectral range from 371-416 nm and 728-758 nm respectively (Figure 1). The Zen operational and data analysis software is very user friendly. The Zeiss Zen software has many functions, including the Multi-Timing Macro, Multicolor+3D+time, FRAP, FRET, tile imaging, multitrack scanning to avoid crosstalk, various image analysis functions (e.g. 3D projection, linear unmixing), etc (see table1).
Coupling the Zeiss 980 with a • Spectra-Physics Insight 3X, ultrafast Ti:Sapphire laser (pre-compensation provides 120 femto second (fs) pulses, digitally tunable range from 680nm to 1300nm, with an average peak power of >1 W) will provide an automated dispersion compensation that delivers shorter pulses at the specimen plane. The combination of short pulses at high power level provides brighter images from deep inside tissue specimens or whole animals. The users have the freedom to select the wavelength from the software instead of looking for an expert to tune the required wavelength and stabilize the laser pulse.
The components equipped with the Zeiss 780 system
- Objective lenses: 5X/0.16, 10X/0.45, 20X/0.80, 40X/1.30 (Oil), 63X/1.40 (Oil),
- Laser lines: 405,445,488,514,561,639
- Spectra-Physics Insight 3X, digitally tunable from 680 to 1300 nm. With dispersion compensation.
- One 8Y Airyscan detector.8 times faster than the normal airyscan mode.
- Internal Detectors: 32-channel GaAsP (Gallium Arsenide Phosphide) spectral detector plus one UV and one Red sensitive PMT detectors.
- External Detectors: Two sensitive Transmission NDD PMT detectors and Two NDD fluorescence GaAsP detectors.
- Automated piezo stage (for screening and tile imaging) and definite focus.
- Temperature controlled Chamber and Eppendorf microinjection device.