Bioimaging Wiki

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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_map_new.png">[Lisbon] 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="http://www.zeiss.de/micro" target="_blank">ZEISS Microscopy</a><br>
<b>Model</b>: <a href="https://www.youtube.com/watch?v=sGTNBXq8omg">LSM 880 with Airyscan</a> (click for Zeiss webinar) <br>
<b>Nickname</b>: "880"<br>
<b>Software</b>: ZEN 2.3 SP1 (black edition) <br>
<b>Year</b>: 2016 <br>
<b>SN</b>: 2811000267 <br>
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Data will be deleted after: <b>1 month</b>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 880 Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Zeiss LSM 880 Usage Statistics</a>
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===== Microscope overview =====

{{  :confocal_pinhole.png?340}} The Zeiss LSM 880 with Airyscan is a [[https://www.youtube.com/watch?v=rTkjQ6Q0p5I|point scanning confocal microscope]] able to generate high-resolution three-dimensional images of thick specimens with high sensitivity, high speed and low photodamage. It is an inverted microscope especially suitable for live cell imaging and photobleaching experiments, equipped with a large cage incubator and a stage top incubator for temperature control and CO2 supply and with Definite Focus for hardware focus control during long time-lapse acquisitions. The stage is motorized and furthermore equipped with a piezo for Z displacement so fast 4D imaging is possible in multiple stage positions. Its Airyscan with 32 concentrically arranged GaAsP detection elements is able to collect all light from an Airy pattern simultaneously, allowing for increased resolution (140 nm laterally and 400 nm axially, at 488 nm). It can also be used in [[https://www.youtube.com/watch?v=ln6VUmLzLLc|Fast Mode]], which enables parallel excitation and detection of four image pixels. The result is a speed improvement by a factor 4x, while maintaining the sensitivity of Airyscan and 1.5x resolution improvement. Its scanning unit further includes two PMT detectors (one of them cooled) as well as a GaAsP detector for increased sensitivity (45% QE compared to ~25% QE for conventional PMT). It is equipped with lasers from violet to far red (405, 458, 488, 514, 561, 594 and 633 nm excitation wavelengths).

  * [[https://www.youtube.com/watch?v=sGTNBXq8omg|ZEISS Webinar: LSM 880 with Airyscan - Revolutionize Your Confocal Imaging]]
  * [[https://www.youtube.com/watch?v=VuxgaKDa_DU|ZEISS Webinar: Fast Mode for ZEISS LSM 880 with Airyscan]]

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<p align="center"><iframe width="420" height="236" src="https://www.youtube.com/embed/QjKrLTHGnc8" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture" allowfullscreen></iframe> <iframe width="420" height="236" src="https://www.youtube.com/embed/ln6VUmLzLLc" title="YouTube video player" frameborder="0" allow="accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture" allowfullscreen></iframe></p>
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{{: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.

===== 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 =====

==== LASERs ====
^  Laser Unit  ^  Wavelength  ^  Maximum Power  ^
|  Diode 405-30  |  405 nm  |  30 mW  |
|  Argon  |  458, 488 and 514 nm  |  25 mW  |
|  DPSS 561-20  |  561 nm  |  20 mW  |
|  HeNe594  |  594 nm  |  2 mW  |
|  HeNe633  |  633 nm  |  5 mW  |

==== Objectives ====
^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  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]]  |
|  20x  |  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]]  |
|  40x  |  C-Apochromat Corr  |  Water  |  1.20  |  0.28  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/421767-9970-000/Objective-C-Apochromat-40x-1.20-W-Corr-M27|421767-9970-000]]  |
|  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]]  |
|  63x  |  Plan-Apochromat DIC  |  Oil  |  1.40  |  0.19  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420782-9900-799/Objective-Plan-Apochromat-63x-1.4-Oil-DIC-M27|420782-9900-799]]  |


**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/420852-9973-000/Objective-LCI-Plan-Neofluar-25x-0.8-Imm-Corr-DIC-M27|420852-9972-000]]  |
|  40x  |  Plan-Apochromat Corr  |  Dry  |  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]]  |
==== Filtersets (Ocular) ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Dichroic  ^  Emission  ^
|  2  |  Cyan  |  [[https://www.chroma.com/products/sets/49001-et-ecfp|49001]]  |  426-446 nm  |  455 nm  |  460-500 nm  |
|  3  |  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  |  495 nm  |  500-550 nm  |
|  4  |  Red  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=000000-1114-101&o=|FS43]]  |  533-558 nm  |  570 nm  |  570-640 nm  |
|  5  |  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]]  |  G 365 nm  |  395 nm  |  420-470 nm  |

===== How to start your session =====

The system should be always on, you only need to:
  * Turn on the HXP lamp
  * Turn on the computer (if needed)
  * Log into Windows with your Agendo credentials
  * Start **ZEN**

===== How to end your session =====

If there is another user for this microscope in the __next hour__:
  * Exit the ZEN software
  * Leave the Ar laser on Standby
  * Leave all other used lasers and HXP lamp on
  * Log off the computer
  * Clean up immersion objectives
Else:
  * Turn off the lasers on **ZEN**
  * Turn off the HXP lamp
  * Clean up immersion objectives
  * Exit the ZEN software
  * Log off the computer

===== Microscope Turn On Procedure (after complete shutdown) =====

If the system is completely off, please follow this procedure:

{{zeiss_lsm_880_1.jpg?100  }}
  * Turn on the ''MAIN SWITCH''
  * Turn on the ''SYSTEMS/PC'' switch
  * Turn on the ''COMPONENTS'' switch
  * Turn on the HXP lamp
  * Turn on the computer and login into Windows with your Agendo credentials
  * Start the **ZEN Black** software

===== Microscope Turn Off Procedure (for complete shutdown) =====

Please only shutdown the system if instructed to do so by the Bioimaging Unit.

  * Turn off the lasers on **ZEN**
  * Exit the ZEN software
  * Shutdown the computer
  * Turn off the HXP lamp
  * Turn off the two switches ''SYSTEMS/PC'' and ''COMPONENTS''
  * **Wait 5 min. for Ar laser to cool down**
  * Turn off the ''MAIN SWITCH'' (**do not turn the key**)

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