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leica_sp8_mp [2020/12/14 12:41]
bioimaging
leica_sp8_mp [2022/09/05 12:18] (current)
bioimaging
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-<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_2022.png"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=leica_sp8_mp_wiki_2022.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">​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<br>
 <​b>​Software version</​b>:​ 3.5.7<​br>​ <​b>​Software version</​b>:​ 3.5.7<​br>​
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 &nbsp <br> &nbsp <br>
 &nbsp <br> &nbsp <br>
-&rarr; <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>​ +&nbsp <br>
- <​br> ​+
 &rarr; <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>​ &rarr; <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). 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|Zeiss 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]]
  
 {{:​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
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 ==== 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 ====
 ^  Magnification ​  ​^ ​ Model  ^  Immersion ​ ^  NA  ^  WD (mm)  ^  Reference ​ ^ ^  Magnification ​  ​^ ​ Model  ^  Immersion ​ ^  NA  ^  WD (mm)  ^  Reference ​ ^
-|  25x  |  HC FLUOTAR VISIR  |  Water  |  0.95  |  2.50  |  15506374 ​ | +|  25x  |  HC FLUOTAR VISIR  |  Water  |  0.95  |  2.50  |  ​[[https://​www.leica-microsystems.com/​objectivefinder/​detail/​objective/​506374/​|15506374]]  | 
-|  63x  |  HC PL APO Glyc CORR CS2  |  Glycerol ​ |  1.30  |  0.30  |  15506353 ​ |+|  63x  |  HC PL APO Glyc CORR CS2  |  Glycerol ​ |  1.30  |  0.30  |  ​[[https://​www.leica-microsystems.com/​objectivefinder/​detail/​objective/​506353/​|15506353]]  |
  
 ==== Filtersets (Ocular) ==== ==== Filtersets (Ocular) ====
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   * Turn on the microscope electronics box   * Turn on the microscope electronics box
 {{::​3_electronics_box.png?​100|}} {{::​3_electronics_box.png?​100|}}
-  * Turn on the microscope'​s ​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
  
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   * 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
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 ---- ----
-[[resources|Back to the Resources ​page]]+[[resources|Back to the Equipment ​page]]
leica_sp8_mp.1607946071.txt.gz · Last modified: 2020/12/14 12:41 by bioimaging