Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=3i_marianas_sdc_wiki_new.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=3i_marianas_sdc_wiki_new.png" width=300></a>
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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=3i_marianas_sdc_map_new.png">Room P2-A-22</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47217) <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</a><br>
<b>Nickname</b>: "Marianas"<br>
<b>Software</b>: SlideBook 6.0.4<br>
<b>Year</b>: 2012 <br>
<b>SN</b>: 1648 <br>
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&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/111"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  3i Marianas SDC Booking</a>
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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 =====


{{  :spinning_disk.png|}} 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. The system is also equipped with a large size incubator for temperature control and a small stage incubator for CO2 supply. With this system you can perform optical sectioning of fluorescent samples which are too thick for a widefield system such as the [[zeiss_cell_observer|Zeiss Cell Observer]]. Even though its resolution is not as high as a //point-scanning confocal system// such as the [[zeiss_lsm_880|Zeiss LSM 880]] or [[zeiss_lsm_710|Zeiss LSM 710]], it is much faster than these two systems. You can also check the [[zeiss_cell_observer_sd|Zeiss Cell Observer SD]] if you need a spinning disk with a Definite Focus system for long time-lapse acquisitions. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. 

  * **Microscope**: [[http://www.zeiss.com/microscopy/en_de/products/light-microscopes/axio-observer-for-biology.html|Zeiss Axio Observer]]
  * **Confocal scanner**: [[https://www.yokogawa.com/eu/solutions/products-platforms/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]]

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

==== LASERs ====
^  Laser Unit  ^  Wavelength  ^  Maximum Power  ^  Current Status  ^
|  Laserstack 405  |  405 nm  |  100 mW  |  ok  |
|  Laserstack 488  |  488 nm  |  100 mW  |  ok  |
|  Laserstack 561  |  561 nm  |  100 mW  |  ok  |
|  Laserstack 640  |  640 nm  |  100 mW  |  ok  |

==== Objectives ====
^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  10x  |  EC Plan-Neofluar  |  Air  |  0.30  |  5.5  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9900-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9900-000]]  |
|  20x  |  Plan-Apochromat  |  Air  |  0.8  |  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]]  |
|  40x  |  LD C-Apochromat Corr  |  Water  |  1.1  |  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]]  |
|  63x  |  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  |  Plan-Apochromat  |  Oil  |  1.40  |  0.17  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420790-9901-000/Objective-Plan-Apochromat-100x-1.40-Oil-M27|420790-9901-000]]  |

**Upon request:**
^  Magnification  ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  25x  |  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]]  |
|  40x  |  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  ====
^  Setting  ^  Transmission  ^
|  1: 445  |  422-477 nm  |
|  2: 525  |  510-540 nm  |
|  3: 617  |  580-653 nm  |
|  4: 692  |  672-712 nm  |
|  5: QUAD  |  440+521+607+700 nm  |
|  6: Block  |  No Transmission  |
|  7: Block  |  No Transmission  |
|  8: Block  |  No Transmission  |
|  9: Empty  |  Empty  |
|  10: 445  |  422-477 nm  |

==== 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 (white socket outlet underneath the touchscreen)
  * Turn on incubation and CO2 on the touchscreen (if needed)
{{marianas_power6.jpg?|}}
  * Turn on the lasers you need to use (using the key and the power switches)
{{marianas_power7.jpg?|}}
  * Turn on the computer
  * Wait 2 min to allow for system components detection
  * Start the **SlideBook** software

**Warning**: If you need to change the stage adaptor, please contact the <html><a href="/facility/bioimaging/doku.php?id=about">Bioimaging Unit</a> (<b>imm-bioimaging@medicina.ulisboa.pt</b> | <img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47222</html>)

===== Microscope Turn Off Procedure =====
  * Remove your sample from the stage/piezo holder
  * Exit the **SlideBook** software
  * Switch off 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 fluorescent lamp
  * Turn off the secondary power supply switch (white socket outlet underneath the touchscreen)
  * Turn off the component power switches (red) **one at a time** from right to left and the power switch distributor (green)

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