leica_sp8_mp
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leica_sp8_mp [2021/08/06 19:17] bioimaging [System Turn On Procedures] |
leica_sp8_mp [2022/02/14 22:15] bioimaging [Microscope overview] |
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | {{ :confocal_vs_2photon.png?0x300}} 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|Zeiss LSM 880]] or the [[zeiss_lsm_710|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|Cell Observer]] or the [[nikon_eclipse_ti|Nikon Eclipse Ti]]. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. | + | {{ :confocal_vs_2photon.png?0x300}} 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|Zeiss LSM 880]] or the [[zeiss_lsm_710|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|Cell Observer]] or the [[nikon_eclipse_ti|Nikon Eclipse Ti]]. If your personal computer cannot handle all the data you collected, check out the [[colossus|Colossus]]. |
| [[https://www.drbio.cornell.edu/cross_sections.html|Two-photon action cross sections - Cornell University]] | [[https://www.drbio.cornell.edu/cross_sections.html|Two-photon action cross sections - Cornell University]] | ||
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| * Turn on the microscope control panel | * Turn on the microscope control panel | ||
| {{::4_control_panel.png?150|}} | {{::4_control_panel.png?150|}} | ||
| - | * Switch on the scanner power, laser power and activate the key switch to turn on the 488 nm laser in the flexible supply unit | + | * Switch on the scanner power, laser power and turn the key switch to on |
| {{::5_scanners_laser.png?120|}} | {{::5_scanners_laser.png?120|}} | ||
| * Log in to Windows (Bioimaging User) | * Log in to Windows (Bioimaging User) | ||
| Line 115: | Line 115: | ||
| * Turn of the 488 nm laser with the key switch in the flexible supply unit | * Turn of the 488 nm laser with the key switch in the flexible supply unit | ||
| * Switch off the HyD supply unit | * Switch off the HyD supply unit | ||
| - | * Switch off the microscope's control panel | + | * Switch off the microscope control panel |
| * Shutdown the computer | * Shutdown the computer | ||
| * Switch off the laser power and scanner power in the flexible supply unit | * Switch off the laser power and scanner power in the flexible supply unit | ||
leica_sp8_mp.txt ยท Last modified: 2024/01/28 12:05 by bioimaging
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