Bioimaging Wiki

The Leica SP8 MP is a both a confocal and multi-photon microscope able to generate high-resolution three-dimensional images of thick specimens. Multi-photon excitation (most commonly two photon excitation) is particularly advantageous for imaging thicker samples. Rather than exciting the fluorophore with one photon, multi-photon excitation is produced by two or more lower energy photons which can penetrate deeper in the sample. Moreover, contrarily to confocal, the multi-photon excitation light will only achieve sufficient intensity to cause fluorescence in a specific region. Because of this, no pinhole is needed in multi-photon microscopy to exclude the light from out-of-focus planes and achieve optical sectioning. The Leica SP8 MP is an upright microscope equipped with water and glycerol immersion objectives especially suitable for intra-vital and thick samples immersed in water or glycerol imaging. In confocal mode, its scanning unit includes a spectral detector PMT to be used with a 488 nm laser for excitation. In multi-photon mode, its Insight DS+ Dual pulsed laser can be tuned from 680 to 1300 nm and has a second laser line emitting at 1041 nm, to be used with four non-descanned detectors with specific filtersets: two PMTS and two HyD detectors (hybrid detectors with 45% QE compared to ~25% QE for conventional PMT). With this system you can perform optical sectioning high resolution imaging of fluorescent samples that are two thick for confocal microscopes such as the Zeiss LSM 880 or the Zeiss LSM 710, albeit with a slightly lower resolution. If you do not need optical sectioning and your sample is thin, check out a widefield system instead, such as the Zeiss Cell Observer or the Nikon Eclipse Ti. If your personal computer cannot handle all the data you collected, check out the Colossus.

Click on the image on the right to see it in higher detail

Data files older than 3 months will be automatically deleted on this system, please copy your data to the iMM server using the desktop link.

Upon request:
(requires changing filters and the dichroic beamsplitter)

• Check that the cooling and power supply for the infrared laser are ON (InSight DeepSee READY)

• Turn on the fluorescence lamp

• Turn on the HyD (Hybrid detectors) supply unit

• Turn on the computer
• Turn on the microscope electronics box

• Turn on the microscope control panel

• Switch on the scanner power, laser power and turn the key switch to on

• Login in to Windows with your Agendo credentials
• Start the LAS X software

If there is another user for this microscope in the next hour:

• Leave the fluorescent lamp and lasers on
• Clean up immersion objectives

Else:

• Clean up immersion objectives
• Set the infrared laser to hibernate (deactivate the infrared laser in the Currently Available Lasers window)
• Close the LAS X software
• Turn of the 488 nm laser with the key switch in the flexible supply unit
• Switch off the HyD supply unit
• Switch off the microscope control panel
• Shutdown the computer
• Switch off the laser power and scanner power in the flexible supply unit
• Turn off the microscope electronics box
• Turn off the fluorescent lamp

Back to the Equipment page