Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_wiki.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_wiki.jpg" width=300></a>
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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_map.jpg">Room P2-A-22</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47217) <br> 
<b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><br>
<b>Model</b>: <a href="http://www.zeiss.com/microscopy/en_de/products/confocal-microscopes/lsm-880-with-airyscan-.html" target="_blank">LSM 880 with Airyscan</a><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://imm.medicina.ulisboa.pt/intranet/booking/new/128"><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.jpg?0x380}} 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 and low photodamage. It is an inverted microscope specially 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. 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). 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). 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_wf|Zeiss Cell Observer WF]] 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 faster confocal systems such as the spinning disk [[3i_marianas_sdc|3i Marianas SDC]] or the //line scanning// [[zeiss_lsm_7_live|Zeiss LSM 7 Live]]. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]].

{{info.gif?|}} Click on the image on the left to see the system beam path in higher detail

===== Booking Rules =====
The Zeiss LSM 710 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)
  - Users must write their internal extension phone number in the booking Description field

===== 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  ^  Type  ^  NA  ^  WD (mm)  ^
|  10x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420340-9901-000|EC Plan-Neofluar]]  |  Dry  |  0.30  |  5.2  |
|  20x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420650-9901-000|Plan-Apochromat]]  |  Dry  |  0.80  |  0.55  |
|  63x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440762-9904-000|Plan-Apochromat]]  |  Oil  |  1.40  |  0.19  |
|  100x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440782-9902-000|Plan-Apochromat]]  |  Oil  |  1.40  |  0.17  |

**Upon request:**

^  Magnification   ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
|  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  |
|  100x  |  [[https://www.micro-shop.zeiss.com/?s=7138707e1a1de&l=en&p=uk&f=o&a=v&m=s&id=420792-9800-000|alpha Plan-Apochromat]]  |  Oil  |  1.46  |  0.11  |

==== Filtersets (Ocular) ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  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  |  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  |  570-640 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  |  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 the computer on
  * Login with your unit account
  * Wait for network icon to change to ''connected'' state
  * Start the **ZEN** software

===== Microscope Turn Off Procedure =====
If there's another user for this microscope in the next hour:
  * Close the software, leave the lasers on and log off the computer. 
  * Clean up any objective where you used immersion oil.
  * **Make sure that there really is another user going to use the microscope.**
Else:
  * Turn off the lasers on **ZEN**
  * Clean up any objective were you used immersion oil.
  * Exit the software, shut down the computer.
  * Turn off the main switches ''SYSTEMS/PC'' and ''COMPONENTS''
  * Wait 5 min. for Ar laser cooldown
  * Turn off the ''MAIN SWITCH''.

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