GIMM Bioimaging Wiki

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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=bartolomeu_dias_wing_2025_-_leica_sdc.png">[Oeiras] Room 0B04</a><br>
<b>Manufacturer</b>: Andor <br>
<b>Model</b>: Revolution XD <br>
<b>Nickname</b>: Leica Spinning Disk<br>
<b>Software</b>: MetaMorph<br>
<b>Year</b>: 2016 <br>
<b>SN</b>: 427435<br>
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Data will be deleted after: <b>3 months</b>
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&rarr; <a href="/facility/bioimaging/doku.php?id=leica_sd_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Leica Spinning Disk Confocal Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=leica_sd_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Leica Spinning Disk Confocal Usage Statistics</a>
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===== Microscope overview =====

{{  :yokogawa_sd.png?350|}} The Leica Spinning Disk Confocal Microscope is a high-performance imaging system designed for dynamic live cell studies, offering a balance between speed, resolution, and confocality. It uses a spinning disk with multiple pinholes to block out-of-focus light, while a sensitive EMCCD Andor [[https://andor.oxinst.com/products/ixon-emccd-camera-series/ixon-ultra-888|iXon Ultra 888]] camera enables rapid acquisition at 1024x1024 resolution. The motorized stage, equipped with a piezo for precise Z-axis control, supports advanced 4D imaging across multiple positions and channels. Adaptive Focus Control corrects vertical drift during long experiments, and the system includes a small cage and stage-top incubators for precise temperature and CO₂ regulation, ensuring stable conditions for fast, sensitive imaging with minimal photobleaching.

  * **Microscope**: [[https://www.leica-microsystems.com/products/light-microscopes/p/dmi8/|Leica DMi8]]
  * **Confocal scanner**: Andor Revolution XD
  * **Camera**: [[https://andor.oxinst.com/products/ixon-emccd-camera-series/ixon-ultra-888|Andor iXon Ultra 888]]

{{:warning.png?25|}} Data files older than **1 month** will be automatically deleted on this system, please copy your data to the GIMM server using the desktop link.

===== System components =====

==== Lasers ====

^  LASER  ^  Wavelength  ^  Maximum Power  ^
|  Violet  |  <fc #a600ff>405 nm</fc>  |  100 mW  |
|  Blue  |  <fc #00f7ff>488 nm</fc>  |  100 mW  |
|  Yellow-Green  |  <fc #c6ff00>561 nm</fc>  |  100 mW  |
|  Red  |  <fc #ff2100>640 nm</fc>  |  100 mW  |

==== Objectives ====

^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Pixel Size (μm/pixel)  ^  Reference  ^
|  <fc #fef042>10x</fc>  |  -  |  Air  |  0.30  |  7.8  |  1.300  |  -  |
|  <fc #0aa05c>20x</fc>  |  HC PL APO  |  Air  |  0.80  |  0.4  |  0.650  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506529/|506529]]  |
|  <fc #2c80c0>63x</fc>  |  HC PL APO  |  Gly  |  1.30  |  0.3  |  0.137  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506398/|506398]]  |
|  100x  |  HC PL APO  |  Oil  |  1.40  |  0.13  |  0.130  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506372/|506372]]  |

==== Emission Filters Spinning Disk ====
 
^  Filter name  ^  Transmission  ^  Most common fluorophores  ^
|  CSU 405 BP  |  <fc #0035ff>417-477 nm</fc>  |  DAPI or Hoechst  |
|  CSU 488 BP  |  <fc #4aff00>500-550 nm</fc>  |  GFP, FITC or Alexa488  |
|  CSU 561 BP  |  <fc #ffa600>589-625 nm</fc>  |  RFP, TRITC or A568  |
|  CSU 640 BP  |  <fc #ff0000>665-705 nm</fc>  |  Cy5, A633 or A647  |
|  DIC  |  -	  |  Bright field with DIC optics  |
|  Quad  |  <fc #0041ff>440</fc>+<fc #4aff00>521</fc>+<fc #ffa600>607</fc>+<fc #ff0000>700</fc> nm  |  DAPI+A488+A568/A647  |


===== Microscope Turn On Procedure =====

  * Turn on the computer (if needed)
  * Turn the power bar in the left side of the table On by pressing the red button, a green light should turn on.
  * Turn the power bar on the right side of the table On. This power bar in on top of the table behind the computer screen.
  * Turn on the Fluorescent lamp if you need it. There are 2 buttons in the lamp: power supply (1) [left side] and LED remote shutter(2) [right side].
  * Turn on the CSU unit (Spinning Disk) by rotating the key to the On position.
  * Turn on the Camera.
  * Open MetaMorph software with the Confocal configuration.

When you are finished and there is someone immediately after you, leave the entire system on. If you're the last or only person of the day, see the turn off procedures below.


===== Microscope Turn Off Procedure =====

  * Close the software and save all your files. Save all the files in the server using the shortcuts provided in the desktop.
  * Turn off the Cameras.
  * Turn off the CSU scanhead (Spinning disk) to the Off position.
  * Turn of the Fluorescent lamp by turning Off the left side button (1), check pictures above in case of doubt.
  * Turn off both power bars (left and right side), check images above in case of doubt.
  * Clean any immersion objectives you used (gently wipe with optic cleaning paper and EtOH embedded paper).
  * TAKE OUT EVERYTHING YOU BROUGHT! (pipettes, slides, etc...)

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