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zeiss_cell_observer_sd [2020/06/01 19:21]
bioimaging
zeiss_cell_observer_sd [2020/10/27 18:28]
bioimaging
Line 10: Line 10:
 <​b>​Model</​b>:​ Cell Observer SD<​br>​ <​b>​Model</​b>:​ Cell Observer SD<​br>​
 <​b>​Nickname</​b>:​ "Cell Observer SD"<​br>​ <​b>​Nickname</​b>:​ "Cell Observer SD"<​br>​
-<​b>​Software</​b>:​ ZEN<​br>​+<​b>​Software</​b>:​ ZEN 2.6 (blue edition)<br>
 <​b>​Year</​b>:​ 2015 <br> <​b>​Year</​b>:​ 2015 <br>
 <​b>​SN</​b>:​ 3851001637 <br> <​b>​SN</​b>:​ 3851001637 <br>
Line 30: Line 30:
 ===== System overview ===== ===== System overview =====
  
-{{  :​spinning_disk.png|}} The Zeiss Cell Observer SD //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, a small stage incubator for CO2 supply and with Definite Focus 2 for hardware focus control during ​long time-lapse acquisitions. You can also use the system as a widefield microscope, using a sCMOS camera for acquisition. Even though its resolution is not as high as a //​point-scanning confocal system// such as the [[zeiss_lsm_710|Zeiss LSM 710]], it is much faster. You can also check the [[3i_marianas_sdc|3i Marianas SDC]] if you need a spinning disk with higher laser power illumination. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]].+{{  :​spinning_disk.png|}} The Zeiss Cell Observer SD //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, a small stage incubator for CO2 supply and with Definite Focus.2 for hardware focus control during ​large tile or multi-position/​time-lapse acquisitions ​(check tutorial videos below). You can also use the system as a widefield microscope, using a sCMOS camera for acquisition. Even though its resolution is not as high as a //​point-scanning confocal system// such as the [[zeiss_lsm_710|Zeiss LSM 710]], it is much faster. You can also check the [[3i_marianas_sdc|3i Marianas SDC]] if you need a spinning disk with higher laser power illumination. 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]]   * **Microscope**:​ [[http://​www.zeiss.com/​microscopy/​en_de/​products/​light-microscopes/​axio-observer-for-biology.html|Zeiss Axio Observer]]
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   * **sCMOS Camera**: [[https://​www.hamamatsu.com/​resources/​pdf/​sys/​SCAS0081E_C11440-22CU.pdf|Hamamatsu ORCA-flash4.0 V2]]   * **sCMOS Camera**: [[https://​www.hamamatsu.com/​resources/​pdf/​sys/​SCAS0081E_C11440-22CU.pdf|Hamamatsu ORCA-flash4.0 V2]]
   * [[https://​www.zeiss.com/​microscopy/​int/​products/​light-microscopes/​axio-observer-for-biology/​definite-focus.html|Definite Focus.2]]   * [[https://​www.zeiss.com/​microscopy/​int/​products/​light-microscopes/​axio-observer-for-biology/​definite-focus.html|Definite Focus.2]]
 +    * | [[https://​www.youtube.com/​watch?​v=Che0bouWcVE|Introduction to Definite Focus.2]]
 +    * | [[https://​www.youtube.com/​watch?​v=I_c3SAHF984|Perform Large Tile Experiments with Definite Focus.2]]
 +    * | [[https://​www.youtube.com/​watch?​v=IUvdvSNsE1I|Multi-Position Experiments with Definite Focus.2]]
 +    * | [[https://​www.youtube.com/​watch?​v=DqqV6cs3mZg|Multi-Position Time Series with Definite Focus.2]]
  
 {{:​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. {{:​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.
  
 +===== Booking Rules =====
 +  - **Users** can book at maximum **8 hours per week**
 +  - This usage restriction does not apply for **weekends** and for working days **before 9:00** and **after 21:00**
 +  - Exceptions to these rules require **approval from [[joserino@medicina.ulisboa.pt|José Rino]]**.
 + 
 ===== System components ===== ===== System components =====
  
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 ^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^ ^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^
 |  Solid State Laser 405  |  405 nm  |  50 mW  |  ok  | |  Solid State Laser 405  |  405 nm  |  50 mW  |  ok  |
-|  Solid State Laser 488  |  488 nm  |  100 mW  |  ​**not working** ​ |+|  Solid State Laser 488  |  488 nm  |  100 mW  |  ​ok  |
 |  Solid State Laser 561  |  561 nm  |  75 mW  |  ok  | |  Solid State Laser 561  |  561 nm  |  75 mW  |  ok  |
 |  Solid State Laser 638  |  638 nm  |  75 mW  |  ok  | |  Solid State Laser 638  |  638 nm  |  75 mW  |  ok  |
Line 63: Line 72:
 |  10x  |  EC Plan-Neofluar ​ |  Air  |  0.30  |  5.20  |  [[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​420340-9901-000/​Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9901-000]] ​ | |  10x  |  EC Plan-Neofluar ​ |  Air  |  0.30  |  5.20  |  [[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​420340-9901-000/​Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9901-000]] ​ |
 |  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]] ​ | |  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]] ​ |
-|  63x  |  Plan-Apochromat Ph3  |  Oil  |  1.40  |  0.19  |  [[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​420781-9910-000/​Objective-Plan-Apochromat-63-1.4-Oil-Ph3-M27|420781-9910-000]] ​ | 
  
 ==== Filtersets (Widefield) ==== ==== Filtersets (Widefield) ====
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 |  :::  |  :::  |  :::  |  620-642 nm  |  650 nm  |  658-704 nm  | |  :::  |  :::  |  :::  |  620-642 nm  |  650 nm  |  658-704 nm  |
 |  6  |  Empty  ||||| |  6  |  Empty  |||||
 +
 ==== Emission Filters (Confocal) ==== ==== Emission Filters (Confocal) ====
  
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 |  [[https://​www.photometrics.com/​wp-content/​uploads/​2019/​10/​Evolve512-Datasheet.pdf|Photometrics Evolve 512]]  |  EMCCD  |  512 x 512  |  16 x 16  |  > 90%  | |  [[https://​www.photometrics.com/​wp-content/​uploads/​2019/​10/​Evolve512-Datasheet.pdf|Photometrics Evolve 512]]  |  EMCCD  |  512 x 512  |  16 x 16  |  > 90%  |
 |  [[https://​www.hamamatsu.com/​resources/​pdf/​sys/​SCAS0081E_C11440-22CU.pdf|Hamamatsu ORCA-flash4.0 V2]]  |  sCMOS  |  2048 x 2048  |  6.5 x 6.5  |  82%  | |  [[https://​www.hamamatsu.com/​resources/​pdf/​sys/​SCAS0081E_C11440-22CU.pdf|Hamamatsu ORCA-flash4.0 V2]]  |  sCMOS  |  2048 x 2048  |  6.5 x 6.5  |  82%  |
 +
 +===== Microscope Turn On Procedure =====
 +  * Turn on the computer
 +  * Login in to Windows (Bioimaging User)
 +  * Turn on the black socket outlets underneath the microscope table by order (1→2→3)
 +  * Wait 2 min to allow for system components detection
 +  * Start the **ZEN Blue** software
 +
 +**Warning**:​ If you need to change the stage adapter, 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 =====
 +If there is another user for this microscope in the __next hour__:
 +  * Exit the **ZEN** software, leave the lasers on
 +  * Log off the computer
 +  * Clean up immersion objectives
 +Else:
 +  * Exit the **ZEN** software
 +  * Shut down the computer
 +  * Clean up immersion objectives
 +  * Turn off the black socket outlets underneath the microscope table by reverse order (3→2→1)
  
 ---- ----
  
 [[resources|Back to the Resources page]] [[resources|Back to the Resources page]]
zeiss_cell_observer_sd.txt · Last modified: 2024/01/28 11:44 by bioimaging