leica_sp8_mp
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leica_sp8_mp [2021/08/06 19:17] bioimaging [System Turn On Procedures] |
leica_sp8_mp [2025/01/14 16:44] (current) bioimaging [Microscope overview] |
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| - | <img src="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_name_new.png"> | + | <img src="/facility/bioimaging/lib/exe/fetch.php?media=lisbon_leica_sp8_mp_name_gimm_2024.png"> |
| <table style="border:0px solid white;"> | <table style="border:0px solid white;"> | ||
| <tr style="border:0px solid white;"> | <tr style="border:0px solid white;"> | ||
| <td style="border:0px solid white;"> | <td style="border:0px solid white;"> | ||
| - | <a href="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_wiki_new.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_wiki_new.png" width=300></a> | + | <a href="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_wiki_2024.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_wiki_2024.png" width=300></a> |
| <td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
| - | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_map_new.png">Room P2-B-42</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47246) <br> | + | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=leica_sp8_mp_map_new.png">[Lisbon] Room P2-B-42</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47236) <br> |
| <b>Manufacturer</b>: <a href="https://www.leica-microsystems.com/" target="_blank">Leica Microsystems</a><br> | <b>Manufacturer</b>: <a href="https://www.leica-microsystems.com/" target="_blank">Leica Microsystems</a><br> | ||
| <b>Model</b>: <a href="https://www.leica-microsystems.com/products/confocal-microscopes/p/leica-tcs-sp8-mp/" target="_blank">TCS SP8 MP</a><br> | <b>Model</b>: <a href="https://www.leica-microsystems.com/products/confocal-microscopes/p/leica-tcs-sp8-mp/" target="_blank">TCS SP8 MP</a><br> | ||
| - | <b>Nickname</b>: "Multiphoton"<br> | + | <b>Nickname</b>: "SP8"<br> |
| - | <b>Software</b>: LAS X<br> | + | <b>Software</b>: LAS X 3.5.7<br> |
| - | <b>Software version</b>: 3.5.7<br> | + | |
| <b>Year</b>: 2017 <br> | <b>Year</b>: 2017 <br> | ||
| <b>SN</b>: 8100001758 <br> | <b>SN</b>: 8100001758 <br> | ||
| + |   <br> | ||
| + | Data will be deleted after: <b>3 months</b> | ||
|   <br> |   <br> | ||
|   <br> |   <br> | ||
| - | → <a href="https://my.agendo.science/calendar/?id=MzRjYWxlbmRhcnM="><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Leica SP8 MP Booking</a> | ||
| - | <br> | ||
| → <a href="/facility/bioimaging/doku.php?id=leica_sp8_mp_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Leica SP8 MP Quality Control</a> | → <a href="/facility/bioimaging/doku.php?id=leica_sp8_mp_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Leica SP8 MP Quality Control</a> | ||
| <br> | <br> | ||
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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). |
| - | + | ||
| - | [[https://www.drbio.cornell.edu/cross_sections.html|Two-photon action cross sections - Cornell University]] | + | |
| {{:info.png?25|}} Click on the image on the right to see it in higher detail | {{:info.png?25|}} Click on the image on the right to see it in higher detail | ||
| - | {{:warning.png?25|}} 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. | + | {{:warning.png?25|}} Data files older than **3 months** will be automatically deleted on this system, please copy your data to the GIMM server using the desktop link. |
| ===== System components ===== | ===== System components ===== | ||
| ==== LASERs ==== | ==== LASERs ==== | ||
| - | ^ Laser Unit ^ Wavelength ^ Maximum Power ^ Current Status ^ | + | ^ Laser Unit ^ Wavelength ^ |
| - | | InSight DS+ Dual (multi-photon) | 680 - 1300 nm | 1.3 W | ok | | + | | InSight DS+ Dual (multi-photon) | 680 - 1300 nm | |
| - | | ::: | 1040 nm | 1.5 W | ok | | + | | ::: | 1040 nm | |
| - | | SS OBIS 488-20 (confocal) | 488 nm | 20 mW | ok | | + | | SS OBIS 488-20 (confocal) | 488 nm | |
| ==== Objectives ==== | ==== Objectives ==== | ||
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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) | + | * Login in to Windows with your Agendo credentials |
| * Start the **LAS X** software | * Start the **LAS X** software | ||
| Line 115: | Line 112: | ||
| * 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.1628270262.txt.gz · Last modified: 2021/08/06 19:17 by bioimaging
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