Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_wiki_new.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_wiki_new.png" width=300></a>
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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_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="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><br>
<b>Model</b>: LSM 880 with Airyscan<br>
<b>Nickname</b>: "880"<br>
<b>Software</b>: ZEN 2.1 (black) <br>
<b>Year</b>: 2016 <br>
<b>SN</b>: 2811000267 <br>
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&rarr; <a href="https://my.agendo.science/calendar/?id=OGNhbGVuZGFycw=="><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss LSM 880 Booking</a>
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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 =====

{{  :airyscan_beampath.png?400}} The Zeiss LSM 880 with Airyscan is a //confocal point-scanning 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 size 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 [[http://www.zeiss.com/content/dam/Microscopy/Products/laser-scan/LSM%20880/zeiss-lsm880-airyscan-principle-672.jpg|Airyscan]] 32 channel area detector 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 of four, 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). 
{{::fast_airyscan.png?420  }}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]] or the [[zeiss_axiovert_200m|Zeiss Axiovert 200M]]. Image resolution with Airyscan is higher than the [[zeiss_lsm_710|Zeiss LSM 710]] and detection sensitivity is also higher than the spinning disks [[3i_marianas_sdc|3i Marianas SDC]] and [[zeiss_cell_observer_sd|Zeiss Cell Observer SD]]. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]].

{{:info.png?25|}} Click [[https://imm.medicina.ulisboa.pt/facility/bioimaging/lib/exe/fetch.php?cache=&media=airyscan.jpg|here]] to see the system beam path in higher detail

{{: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 =====
The Zeiss LSM 880 booking is divided in three daily slots:
|  9:00 to 13:00  |  13:00 to 17:00  |  17:00 to 21:00  |

  - A **user** can book at maximum **2 sessions per week**
  - Each **session must fit inside one of the three daily slots** (i.e. a session booked from 11:00 to 14:00 counts as 2 sessions)
  - The above rules do not apply for **weekends** and for work days **before 9:00** and **after 21:00**
  - The above rules do not apply for sessions booked on the very same day or **24 hours ahead**
  - Exceptions to these rules require **approval from [[joserino@medicina.ulisboa.pt|José Rino]]**.
  - Users must indicate which lasers they are going to use in the booking Description field
  - Users must indicate if they will use the incubator in the booking Description field (write down 37C INCUBATOR)

===== System components =====

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

==== 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]]  |
|  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]]  |
|  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]]  |
==== Filtersets (Ocular) ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Dichroic  ^  Emission  ^
|  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  |  495 nm  |  500-550 nm  |
|  2  |  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  |
|  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]]  |  G 365 nm  |  395 nm  |  420-470 nm  |

===== Microscope Turn On Procedure =====
  * Turn on the ''MAIN SWITCH''
  * Turn on the two switches: ''SYSTEMS/PC'' and ''COMPONENTS''
{{zeiss_lsm_880_1.jpg?100}}
  * Turn on the computer
  * Login in to Windows (Bioimaging User)
  * Start the **ZEN Black** software

===== 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:
  * Turn off the lasers on **ZEN**
  * Clean up immersion objectives
  * Exit the ZEN software
  * Shutdown the computer
  * Turn off the two switches ''SYSTEMS/PC'' and ''COMPONENTS''
  * **Wait 5 min. for Ar laser to cool down**
  * Turn off the ''MAIN SWITCH''

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