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leica_sp8_mp [2020/01/17 13:46] bioimaging [Emission Filters (multi-photon)] |
leica_sp8_mp [2021/01/17 04:44] bioimaging [Microscope Turn Off procedures] |
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<b>Nickname</b>: "Multiphoton"<br> | <b>Nickname</b>: "Multiphoton"<br> | ||
<b>Software</b>: LAS X<br> | <b>Software</b>: LAS X<br> | ||
- | <b>Software version</b>: 3.5.5.19976<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> |   <br> | ||
  <br> |   <br> | ||
- | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/146"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Leica SP8 MP Booking</a> | + | → <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> | <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> | ||
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==== LASERs ==== | ==== LASERs ==== | ||
^ Laser Unit ^ Wavelength ^ Maximum Power ^ Current Status ^ | ^ Laser Unit ^ Wavelength ^ Maximum Power ^ Current Status ^ | ||
- | | InSight DS+ Dual (multi-photon) | 680 - 1300 nm | 1.3 W | **not working** | | + | | InSight DS+ Dual (multi-photon) | 680 - 1300 nm | 1.3 W | ok | |
- | | ::: | 1041 nm | 1.5 W | **not working** | | + | | ::: | 1040 nm | 1.5 W | ok | |
| SS OBIS 488-20 (confocal) | 488 nm | 20 mW | ok | | | SS OBIS 488-20 (confocal) | 488 nm | 20 mW | ok | | ||
==== Objectives ==== | ==== Objectives ==== | ||
^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
- | | 25x | HC FLUOTAR VISIR | Water | 0.95 | 2.5 | 15506374 | | + | | 25x | HC FLUOTAR VISIR | Water | 0.95 | 2.50 | 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 | 15506353 | | ||
==== Filtersets (Ocular) ==== | ==== Filtersets (Ocular) ==== | ||
- | ^ Filterset ^ Reference ^ Excitation ^ Emission ^ Fluorochromes ^ | + | ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ Fluorochromes ^ |
- | | Blue | I3 | 450-490 nm | > 515 nm | GFP, FITC, Alexa488 | | + | | Blue | I3 | 450-490 nm | 510 nm | > 515 nm | GFP, FITC, Alexa488 | |
==== Emission Filters (multi-photon) ==== | ==== Emission Filters (multi-photon) ==== | ||
+ | {{::default_configuration.png |}} | ||
+ | \\ | ||
+ | \\ | ||
+ | \\ | ||
^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ||
| HyD-RLD 1 | Red | 570-640 nm | mCherry, Alexa568, Cy3 | | | HyD-RLD 1 | Red | 570-640 nm | mCherry, Alexa568, Cy3 | | ||
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| PMT-RLD 3 | Far Red | 662 - 737 nm | Alexa647, Cy5 | | | PMT-RLD 3 | Far Red | 662 - 737 nm | Alexa647, Cy5 | | ||
| PMR-RLD 4 | Red | 570-640 nm | mCherry, Alexa568, Cy3 | | | PMR-RLD 4 | Red | 570-640 nm | mCherry, Alexa568, Cy3 | | ||
+ | \\ | ||
+ | \\ | ||
+ | \\ | ||
+ | \\ | ||
+ | \\ | ||
**Upon request:** | **Upon request:** | ||
- | (requires changing the dichroic beamsplitter) | + | \\ (requires changing filters and the dichroic beamsplitter) |
+ | {{::alternative_configuration.png |}} | ||
+ | \\ | ||
+ | \\ | ||
+ | \\ | ||
^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ^ Detector ^ Filter ^ Transmission ^ Fluorochromes ^ | ||
| HyD-RLD 1 | Green | 500-550 nm | GFP, Alexa488, FITC | | | HyD-RLD 1 | Green | 500-550 nm | GFP, Alexa488, FITC | | ||
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| PMT-RLD 3 | Far Red | 662 - 737 nm | Alexa647, Cy5 | | | PMT-RLD 3 | Far Red | 662 - 737 nm | Alexa647, Cy5 | | ||
| PMR-RLD 4 | Red | 570-640 nm | mCherry, Alexa568, Cy3 | | | PMR-RLD 4 | Red | 570-640 nm | mCherry, Alexa568, Cy3 | | ||
+ | \\ | ||
+ | \\ | ||
+ | \\ | ||
+ | |||
+ | ===== System Turn On Procedures ===== | ||
+ | * Check that the cooling and power supply for the infrared laser are ON (''InSight DeepSee READY'') | ||
+ | {{::0_infrared_laser.png?100|}} | ||
+ | * Turn on the fluorescence lamp | ||
+ | {{1_metal_halide.png?100|}} | ||
+ | * Turn on the HyD (Hybrid detectors) supply unit | ||
+ | {{::2_hyd_supply.png?100|}} | ||
+ | * Turn on the computer | ||
+ | * Turn on the microscope electronics box | ||
+ | {{::3_electronics_box.png?100|}} | ||
+ | * Turn on the microscope's control panel | ||
+ | {{::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 | ||
+ | {{::5_scanners_laser.png?120|}} | ||
+ | * Log in to Windows (Bioimaging User) | ||
+ | * Start the **LAS X** software | ||
+ | |||
+ | ===== Microscope Turn Off procedures ===== | ||
+ | |||
+ | 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's 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 | ||
---- | ---- | ||
- | [[resources|Back to the Resources page]] | + | [[resources|Back to the Equipment page]] |