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zeiss_cell_observer_sd [2016/07/01 11:15]
bioimaging created
zeiss_cell_observer_sd [2020/02/26 11:45]
bioimaging [Objectives]
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 <​html>​ <​html>​
-<img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=3i_marianas_sdc_name.jpg">+<img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_cell_observer_sd_name_new.png">
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-<a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=3i_marianas_sdc_wiki.jpg"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=3i_marianas_sdc_wiki.jpg" width=300></​a>​+<a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_cell_observer_sd_wiki_new.png"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_cell_observer_sd_wiki_new.png" width=300></​a>​
 <td style="​border:​0px solid white;">​ <p style="​line-height:​1.8">​ <td style="​border:​0px solid white;">​ <p style="​line-height:​1.8">​
-<​b>​Location</​b>:​ <a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=3i_marianas_sdc_map.jpg">​Room P2-A-22</​a>​ (<img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=telephone_icon.gif" width=15>​ 47217) <br>  +<​b>​Location</​b>:​ <a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_cell_observer_sd_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>​Manufacturer</​b>:​ <a href="http://www.zeiss.de/micro" target="​_blank">​Carl Zeiss Microimaging</​a><​br>​ 
-<​b>​Model</​b>:​ <a href="​https://​www.intelligent-imaging.com/​marianas.php"​ target="​_blank"​>Marianas</a><​br>​ +<​b>​Model</​b>: ​Cell Observer SD<br> 
-<​b>​Software</​b>: ​SlideBook 6.0.4<​br>​ +<b>Nickname</b>: "Cell Observer SD"<​br>​ 
-<​b>​Year</​b>: ​2012 <​br>​ +<​b>​Software</​b>: ​ZEN<​br>​ 
-<​b>​SN</​b>: ​1648 <br>+<​b>​Year</​b>: ​2015 <​br>​ 
 +<​b>​SN</​b>: ​3851001637 ​<br>
 &nbsp <br> &nbsp <br>
 &nbsp <br> &nbsp <br>
 &nbsp <br> &nbsp <br>
-&nbsp <​br>​ +&rarr; <a href="​https://​imm.medicina.ulisboa.pt/​intranet/​booking/​119"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=date.png">  ​Zeiss Cell Observer SD Booking</​a>​
-&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>​+
  <​br> ​  <​br> ​
-&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>​+&rarr; <a href="/​facility/​bioimaging/​doku.php?​id=zeiss_cell_observer_sd_quality"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=list-icon.png"​ width=18> ​Zeiss Cell Observer SD Quality Control</​a>​
  <​br>​  <​br>​
-&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>​+&rarr; <a href="/​facility/​bioimaging/​doku.php?​id=zeiss_cell_observer_sd_usage"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=chart_line.png">  ​Zeiss Cell Observer SD Usage Statistics</​a>​
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 ===== 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 controla small stage incubator for CO2 supply ​and with Definite Focus 2 for hardware focus control during long time-lapse acquisitionsYou 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_880|Zeiss LSM 880]] or [[zeiss_lsm_710|Zeiss LSM 710]], it is much faster than these two systems. ​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.jpg?0x300|}} 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_axio_observer|Zeiss Axio 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. ​If you need to use an upright microscope, ​check the [[zeiss_lsm_7_live|Zeiss LSM 7 Live]] fast confocal system instead. 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]]
-  * **Confocal scanner**: [[http://www.andor.com/microscopy_systems/peripherals/confocal_scanners/|Yokogawa CSU-x1]] +  * **Confocal scanner**: [[https://www.yokogawa.com/eu/solutions/products-platforms/​life-science/​spinning-disk-confocal/​csu-x1-confocal-scanner-unit/|Yokogawa CSU-x1]] 
-  * **Camera**: [[http://​www.photometrics.com/​products/emccdcams/evolve/512.php|Evolve 512 EMCCD]]+  * **EMCCD Camera**: [[https://​www.photometrics.com/​wp-content/uploads/2019/10/​Evolve512-Datasheet.pdf|Evolve 512 EMCCD]] 
 +  * **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]] 
 + 
 +{{:​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 ===== ===== System components =====
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 ==== LASERs ==== ==== LASERs ====
 ^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^ ^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^
-|  ​Laserstack ​488  |  488 nm  |  100 mW  |  ok  | +|  ​Solid State Laser 405  |  405 nm  |  50 mW  |  ok  | 
-|  ​Laserstack ​561  |  561 nm  |  ​100 mW  |  ok  | +|  Solid State Laser 488  |  488 nm  |  100 mW  |  ok  | 
-|  ​Laserstack 640  ​|  ​640 nm  |  ​100 mW  |  ok  |+|  ​Solid State Laser 561  |  561 nm  |  ​75 mW  |  ok  | 
 +|  ​Solid State Laser 638  ​|  ​638 nm  |  ​75 mW  |  ok  |
  
 ==== Objectives ==== ==== Objectives ====
-^  Magnification ​  ​^ ​ Model  ^  Type  ^  NA  ^  WD (mm)  ^ 
-|  20x  |  [[https://​www.micro-shop.zeiss.com/?​s=129353259d52f12&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420650-9901-000|Plan-Apochromat]] ​ |  Dry  |  0.8  |  0.55  | 
-|  63x  |  [[https://​www.micro-shop.zeiss.com/?​s=3407786617d462&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420782-9900-000|Plan-Apochromat]] ​ |  Oil  |  1.40  |  0.19  | 
-|  100x  |  [[https://​www.micro-shop.zeiss.com/?​s=129353259d52f12&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420790-9901-000|Plan-Apochromat]] ​ |  Oil  |  1.40  |  0.17  | 
  
-**Upon request:​** +^  Magnification ​  ​^  Model  ^  ​Immersion ​ ​^ ​ NA  ^  WD (mm)  ​^ ​ Reference ​ ^ 
-^  Magnification ​ ^  Model  ^  ​Type  ​^ ​ NA  ^  WD (mm)  ^ +|  ​10x  |  EC Plan-Neofluar Ph1  |  Air  |  0.3  |  5.20  ​| ​ [[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​420341-9911-000/​Objective-EC-Plan-Neofluar-10x-0.30-Ph1-M27|420341-9911-000]] 
-|  ​25x  ​| ​ [[https://​www.micro-shop.zeiss.com/​?​s=129353259d52f12&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420852-9972-000|LCI Plan-NeoFluar]]  |  ​Oil/​Glyc/​W ​ ​| ​ 0.8  |  0.21  | +|  20x  ​|  ​Plan-Apochromat Ph2  |  Air  ​| ​ 0.8  |  0.55  ​| ​ [[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​420651-9911-000/Objective-Plan-Apochromat-20x-0.8-Ph2-M27|420651-9911-000]]  ​
-|  40x  ​| ​ [[https://​www.micro-shop.zeiss.com/​?​s=129353259d52f12&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420660-9970-000|Plan-Apochromat]] ​ |  ​Dry  ​|  ​0.95  ​|  ​0.25  | +|  ​40x  ​|  ​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]] ​ | 
-==== Emission Filtersets ==== +|  ​40x  ​|  ​Plan-Apochromat DIC  ​| ​ ​Oil  ​|  ​1.40  ​|  ​0.13  |  [[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​420762-9900-000/​Objective-Plan-Apochromat-40x-1.4-Oil-DIC-M27|420762-9900-000]] ​ | 
- Setting ​  Transmission ​ +|  ​63x  ​|  ​Plan-Apochromat DIC  ​| ​ ​Oil  ​|  ​1.40  ​|  ​0.19  ​|  ​[[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​420782-9900-000/​Objective-Plan-Apochromat-63x-1.4-Oil-DIC-M27|420782-9900-000]] ​ | 
-|  ​1445  ​ 422-477 nm  | +|  ​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]] ​ | 
-|  ​2: 525  ​|  ​510-540 nm  | +|  ​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]] ​ | 
-|  ​3: 617  ​|  ​580-653 nm  | + 
-|  ​4: 692  ​|  ​672-712 nm  | +==== Filtersets (Widefield) ====
-|  ​5: QUAD  ​|  ​440+521+607+700 nm  +
-|  ​6Block  ​ No Transmission ​ | +
-|  ​7: Block  ​|  ​No Transmission ​ | +
-|  ​8: Block  ​|  ​No Transmission ​ | +
-|  ​9: Empty  ​|  ​Empty  | +
-|  ​10: 445  ​|  ​422-477 nm  |+
  
-===== Microscope Turn On Procedure ===== +^  Position ​ ^  Filterset ​ ^  Reference ​ ^  Excitation ​ ^  Emission ​ ^ 
-  * Check that the main power supply switch is on (should be on by default) +|  1  |  Green  |  [[https://​www.micro-shop.zeiss.com/?​s=7139069fdc79c&​l=en&p=us&f=f&a=v&b=f&id=489038-9901-000&​o=|FS38HE]] ​ ​|  ​450-490 nm  ​|  ​500-550 nm  | 
-{{marianas_power1.jpg?​|}} +|  2  |  Red  |  [[https://​www.micro-shop.zeiss.com/​?s=4237481102cf7b&​l=en&​p=us&​f=f&​a=v&​b=f&​id=489043-9901-000&​o=|FS43HE]] ​ |  538-562 nm  |  570-640 nm  | 
-  * Turn on the fluorescence lamp (if needed) +|  3  |  Blue  |  [[https://​www.micro-shop.zeiss.com/​?s=163176628fcf844&​l=en&​p=us&​f=f&​a=v&​b=f&​id=488049-9901-000&​o=|FS49]] ​ |  335-383 nm  |  420-470 nm  | 
-{{marianas_power5.jpg?​|}} +|  4  |  Far Red  |  [[https://​www.micro-shop.zeiss.com/​?s=4237481102cf7b&​l=en&​p=us&​f=f&​a=v&​b=f&​id=488050-9901-000&​o=|FS50]] ​ |  625-655 nm  |  665-715 nm  | 
-  * Turn on the secondary power supply switch +|  5  |  QUAD  |  [[https://​www.micro-shop.zeiss.com/?s=4237481102cf7b&​l=en&​p=us&​f=f&​a=v&​b=f&​id=489081-9010-000&​o=|  ​FS81HE]] ​ |  357-393 nm  |  407-412 nm  | 
-{{marianas_power2.jpg?|}} + ::: ​ |  :::  |  :::  |  473-496 nm  |  511-528 nm  | 
-  * Turn on the power switch distributor (green) and the component power switches (red) **one at a time** + :::  |  :::  |  :::  |  545-562 nm  |  577-602 nm  | 
-{{marianas_power3.jpg?|}} + :::  |  :::  |  :::  |  620-642 nm  |  658-704 nm  | 
-  * Turn on the Yokogawa spinning disk scanner (using the key) + 6  |  none  |  |  |  | 
-{{marianas_power4.jpg?|}} +==== Emission Filters (Confocal) ====
-  * 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 Facility</​a>​ (<​b>​imm-bioimaging@medicina.ulisboa.pt<​/b> <img src="/facility/​bioimaging/​lib/​exe/​fetch.php?​media=telephone_icon.gif"​ width=15>​ 47316</​html>​)+^  Position ​ ^  Transmission ​ ^ 
 +|  1BP 450/50  |  425-475 nm  | 
 +|  2: TBP 526 601 688  |  510-542 nm  | 
 +|  :::  |  586-616 nm  |   
 +|  :::  |  663-713 nm  | 
 +|  3: none  |  | 
 +|  4: FE01-520/35  ​ ​502-537 nm  | 
 +|  5: BP 600 50  |  575-625 nm  | 
 +|  6: BP 690/50  |  665-715 nm  |
  
-===== Microscope Turn Off Procedure ===== +==== Cameras ​==== 
-  ​* Remove your sample from the stage/piezo holder + Model   Type  ^  Frame Size  ^  Pixel Size (µm ​^ ​ Quantum Efficiency ​ ^ 
-  * Exit the **SlideBook** software +|  [[https://​www.photometrics.com/​wp-content/​uploads/​2019/​10/​Evolve512-Datasheet.pdf|Photometrics Evolve 512]]  |  EMCCD  |  512 x 512  |  16 x 16  |  > 90%  | 
-  * Switch off the computer +|  [[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%  |
-  * Turn off all system components ​(follow the Turn On Procedure in reverse)+
  
 ---- ----
  
 [[resources|Back to the Resources page]] [[resources|Back to the Resources page]]
zeiss_cell_observer_sd.txt · Last modified: 2024/04/01 23:53 by bioimaging