Confocal · Leica
A flexible confocal platform with 6 laser lines and 4 independent detectors. The SP8’s highly configurable detection system makes it well suited for experiments requiring broad spectral coverage and simultaneous multi-channel acquisition.
Instrument Features
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6 Laser Lines
Broad excitation coverage with 6 laser lines spanning UV to far-red (405, 458, 488, 514, 561, 633 nm). AOTF (Acousto-Optical Tunable Filter) provides precise, electronically controlled laser power adjustment for each line independently.
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4 Independent Detectors
Combination of PMT and HyD (Hybrid Detector) detectors allow simultaneous 4-channel acquisition. HyD detectors offer superior sensitivity and signal-to-noise for dim samples, with photon counting capability.
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Spectral Detection Flexibility
Continuously tunable detection windows on each detector allow precise spectral band selection without changing filters. Minimizes crosstalk between channels and maximizes signal collection for each fluorophore.
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Simultaneous Multi-Channel Acquisition
All 4 channels can be acquired simultaneously, enabling fast multi-color imaging with minimal photobleaching and no time delay between channel acquisitions — critical for co-localization and dynamic studies.
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FRET Capability
Sensitized emission and acceptor photobleaching FRET protocols are well supported. The precise spectral control and multiple laser lines make the SP8 a strong platform for protein-protein interaction studies.
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LAS X Software
Leica Application Suite X provides an intuitive workflow for experiment design, acquisition, and analysis. Includes automated wizards for Z-stacks, tile scans, FRET, and time series with straightforward channel configuration.
Startup Guide
Power On the System
Turn on the main system power switch and the laser launch box. Power on the PC. Allow 20–30 minutes for full initialization and laser warm-up.
Launch LAS X Software
Open Leica Application Suite X (LAS X) from the desktop. The software will auto-detect all hardware components. Confirm all lasers and detectors are listed in the configuration panel.
Activate Laser Lines
In the laser control panel, activate the required laser lines for your fluorophores. Use the AOTF to set initial laser power (start at 5–10%).
Select Objective
Choose your objective from the nosepiece selector in LAS X. Apply immersion oil or water as appropriate for your objective type.
Load Sample & Navigate
Mount your slide on the stage. Switch to Live mode to find your cells and navigate to your region of interest using the joystick or software stage control.
Configure Detection Channels
Set the detection window for each channel by dragging the spectral sliders in the channel configuration panel. Assign each channel to a PMT or HyD detector. Enable simultaneous acquisition for co-localization experiments.
Optimize Signal
Adjust laser power via AOTF and detector gain for each channel. Use the saturation indicator to avoid overexposure. Set pinhole to 1 Airy Unit for optimal optical sectioning.
Set Acquisition & Acquire
Configure Z-stack, tile, or time-lapse settings as needed. Start acquisition and monitor for drift or photobleaching. Save your .lif file to your data folder.
Shut Down
Deactivate all lasers, remove sample, clean the objective, close LAS X, and follow the posted shutdown procedure.
💡 HyD detectors provide the best sensitivity for dim samples — use them for your weakest channels and reserve PMTs for bright channels to get the most out of simultaneous 4-channel acquisition.