GIMM Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=oeiras_marianas_tirf.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=oeiras_marianas_tirf.jpg" width=300></a>
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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=bartolomeu_dias_wing_2025_-_3i_marianas_sdc_tirf_.png">[Oeiras] Room 0B03</a><br> 
<b>Manufacturer</b>: <a href="https://www.intelligent-imaging.com/" target="_blank">Intelligent Imaging Innovations</a><br>
<b>Model</b>: <a href="https://www.intelligent-imaging.com/marianas" target="_blank">Marianas SDC</a><br>
<b>Nickname</b>: "Marianas TIRF"<br>
<b>Software</b>: SlideBook 2026.1<br>
<b>Year</b>: 2012<br>
<b>SN</b>: CS-01648<br>
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Data will be deleted after: <b>15 days</b>
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&rarr; <a href="/facility/bioimaging/doku.php?id=3i_marianas_sdc_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> 3i Marianas SDC Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=3i_marianas_sdc_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  3i Marianas SDC Usage Statistics</a>
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===== System overview =====

{{  :yokogawa_sd.png?350|}} The 3i Marianas SDC //spinning disk confocal microscope// is a fast imaging system which provides a trade-off between confocality, resolution and speed. It is an inverted microscope ideal for live cell applications which require fast acquisition speeds rather than high resolution images. The scanning unit achieves confocality by directing light through a spinning disk with many small pinholes. Images are then acquired with a sensitive EMCCD which allows for very small exposure times but is limited in resolution to 512x512 pixels. The stage is motorized and furthermore equipped with a piezo for Z displacement so fast 4D imaging is possible in multiple stage positions, with a large cage incubator and a stage top incubator for temperature control and CO2 supply allowing for live-cell imaging.
The system is also equipped with a Total Internal Reflection Fluorescence (TIRF) and photomanipulation module, providing motorized TIRF and HILO illumination for selective excitation within ~100–150 nm of the coverslip and integrated photomanipulation capabilities, including FRAP, photoactivation and photoconversion, ideal for studying membrane dynamics, receptor and vesicle movement.

  * **Microscope**: [[https://www.zeiss.com/microscopy/en/products/light-microscopes/widefield-microscopes/axio-observer-for-life-science-research.html|Zeiss Axio Observer]]
  * **Confocal scanner**: [[https://www.yokogawa.com/eu/solutions/products-and-services/life-science/spinning-disk-confocal/csu-x1-confocal-scanner-unit/|Yokogawa CSU-X1]]
  * **Camera**: [[https://www.photometrics.com/wp-content/uploads/2019/10/Evolve512-Datasheet.pdf|Photometrics Evolve 512 EMCCD]]
  * **Features**
    * | [[https://www.intelligent-imaging.com/tirf|Vector3 TIRF and Photomanipulation]]

{{:warning.png?25|}} Data files older than **15 days** 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 #ff4200>640 nm</fc>  |  100 mW  |

==== Objectives ====
^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  <fc #fef042>10x</fc>  |  EC Plan-Neofluar  |  Air  |  0.30  |  5.50  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9900-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9900-000]]  |
|  <fc #0aa05c>20x</fc>  |  Plan-Apochromat  |  Air  |  0.80  |  0.55  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420650-9901-000/Objective-Plan-Apochromat-20x-0.8-M27|420650-9901-000]]  |
|  <fc #01b3f5>40x</fc>  |  LD C-Apochromat Corr  |  Water  |  1.10  |  0.62  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/421867-9970-000/Objective-LD-C-Apochromat-40x-1.1-W-Corr-M27|421867-9970-000]]  |
|  <fc #2c80c0>63x</fc>  |  Plan-Apochromat  |  Oil  |  1.40  |  0.19  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420780-9900-000/Objective-Plan-Apochromat-63x-1.4-Oil-M27|420780-9900-000]]  |
|  100x  |  α Plan-Apochromat DIC  |  Oil  |  1.46  |  0.11  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420792-9800-000/Objective-alpha-Plan-Apochromat-100x-1.46-Oil-DIC-M27|420792-9800-000]]  |

**Upon request:**
^  Magnification  ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  <fc #0aa05c>25x</fc>  |  LCI Plan-Neofluar Corr DIC  |  Oil/Glyc/W  |  0.80  |  0.21  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420852-9972-000/Objective-LCI-Plan-Neofluar-25x-0.8-Imm-Corr-DIC-M27|420852-9972-000]]  |
|  <fc #01b3f5>40x</fc>  |  Plan-Apochromat Corr  |  Air  |  0.95  |  0.25  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420660-9970-000/Objective-Plan-Apochromat-40x-0.95-Corr-M27|420660-9970-000]]  |

==== Emission Filters CSU X1 ====

^  Position  ^  Setting  ^  Transmission  ^
|  1  |  445  |  <fc #0041ff>422-477 nm</fc>  |
|  2  |  525  |  <fc #4aff00>510-540 nm</fc>  |
|  3  |  617  |  <fc #ff8200>580-653 nm</fc>  |
|  4  |  692  |  <fc #ff0000>672-712 nm</fc>  |
|  5  |  QUAD  |  <fc #0041ff>440</fc>+<fc #4aff00>521</fc>+<fc #ffa600>607</fc>+<fc #ff0000>700</fc> nm  |
|  6  |  Block  |  No Transmission  |
|  7  |  Block  |  No Transmission  |
|  8  |  Block  |  No Transmission  |
|  9  |  Empty  |  Empty  |
|  10  |  445  |  <fc #0041ff>422-477 nm</fc>  |

==== Camera ====
^  Model  ^  Frame Size  ^  Pixel Size (µm)  ^  Quantum Efficiency  ^
|  [[https://www.photometrics.com/wp-content/uploads/2019/10/Evolve512-Datasheet.pdf|Photometrics Evolve 512 EMCCD]]  |  512 x 512  |  16 x 16  |  > 90%  |

===== Microscope Turn On Procedure =====
  * Turn on the fluorescence lamp (if needed)
{{marianas_power5.jpg?|}}
  * Turn on the power switch distributor (green) and the component power switches (red) **one at a time** from left to right
{{marianas_power3.jpg?|}}
  * Turn on the secondary power supply switch (socket outlet below the touchscreen)
{{::marianas_power2.png|}}
  * Turn on incubation and CO2 in the touchscreen (if needed)
{{marianas_power6.jpg?|}}
  * Turn on the lasers you need to use (using the key and the power switches)
{{::marianas_power7.png?160|}}
  * Turn on the computer
  * Login in to Windows with your Agendo credentials
  * Wait 2 min to allow for system components detection
  * Start the **SlideBook** software

===== Microscope Turn Off Procedure =====
If there is another user for this microscope in the __next hour__:
  * Exit the **SlideBook** software, leave the lasers on
  * Log off the computer
  * Clean up immersion objectives
Else:
  * Exit the **SlideBook** software
  * Shutdown the computer
  * Turn off incubation in the touchscreen (if used)
  * Turn off CO2 (if used)
  * Switch all lasers off (first on the switch and finally by turning the key)
  * Turn off the secondary power supply switch (white socket outlet below the touchscreen)
  * Turn off the component power switches (red) **one at a time** from right to left and the power switch distributor (green)
  * Turn off the fluorescent lamp

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