Bioimaging Wiki

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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_710_map_new.png">Room P3-A-10</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47331) <br> 
<b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><br>
<b>Model</b>: LSM 710<br>
<b>Nickname</b>: "710"<br>
<b>Software</b>: ZEN 2012<br>
<b>Year</b>: 2010 <br>
<b>SN</b>: 2503000400 <br>
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&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/51"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss LSM 710 Booking</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_710_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 710 Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_710_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Zeiss LSM 710 Usage Statistics</a>
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===== Microscope overview =====
{{  :lsm710_beampath.png?410|}} The Zeiss LSM 710 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 (but no stage incubator for CO2 supply). Its scanning unit has a spectral recycling loop, two sensitive PMT detectors and a filter-free spectral separation module that can be continuously set over the entire wavelength range (i.e. you can specify which wavelength range you want to detect). It is equipped with lasers from violet to far red (405, 458, 488, 514, 561 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|Zeiss Cell Observer]] or the [[zeiss_axiovert_200m|Zeiss Axiovert 200M]]. Image resolution and detection sensitivity are higher than faster confocal systems such as the spinning disks [[3i_marianas_sdc|3i Marianas SDC]] and [[zeiss_cell_observer_sd|Zeiss Cell Observer SD]], but image acquisition is slower. If you need to use a 594 nm laser or increased resolution and sensitivity, or are performing long time-lapse experiments in live samples (aqueous media) and need hardware focus control, check the [[zeiss_lsm_880|Zeiss LSM 880]] confocal point-scanning system with Airyscan and Definite Focus. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]].

{{:warning.png?25|}} Data files older than **3 months** will be automatically deleted on this system, please copy your data to the iMM server using the desktop link.

===== 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)
===== 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 561-10  |  561 nm  |  15 mW  |  ok  |
|  HeNe633  |  633 nm  |  5 mW  |  ok  |

==== Objectives ====
^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  10x  |  EC Plan-Neofluar  |  Air  |  0.30  |  5.2  |  [[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  |  EC Plan-Neofluar DIC  |  Oil  |  1.30  |  0.20  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440452-9903-000/Objective-EC-Plan-Neofluar-40x-1.30-Oil-DIC|440452-9903-000]]  |
|  63x  |  Plan-Apochromat DIC  |  Oil  |  1.40  |  0.19  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440762-9904-000/Objective-Plan-Apochromat-63x-1.40-Oil-DIC|440762-9904-000]]  |

**Upon request:**

^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  100x  |  Plan-Apochromat DIC  |  Oil  |  1.40  |  0.17  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440782-9902-000/Objective-Plan-Apochromat-100x-1.40-Oil-DIC|440782-9902-000]]  |
==== Filtersets (Ocular) ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Dichroic  ^  Emission  ^
|  1  |  Green  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=000000-1031-346&o=|FS38]]  |  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''
{{switches_710.jpg?100}}
  * Turn on the metal halide fluorescent light source (if needed)
{{halide_710.jpg?80}}
  * Turn on the Ar laser using the key (if needed)
{{laser_key_710.jpg?100}}
  * Turn on the computer
  * Log in to Windows (Bioimaging User)
  * Login with Bioimaging User account
  * Check that the computer is connected to an ''Unidentified network'' (click on the network icon in the taskbar)
{{::network_icon.png?200}}
  * Switch the Ar laser from ''idle'' ton ''run'' (don't change the ''current'')
{{laser_run_710.jpg?100}}
  * Start the **ZEN Black** software

===== Microscope Turn Off Procedure =====

If there is another user for this microscope in the __next hour__:
  * Close **ZEN**, leave the lasers on, log off the computer
  * Clean up immersion objectives
Else:
  * Turn off the 561 nm laser and incubator in the software (if used)
  * Switch the Ar laser from ''run'' to ''idle'' (if used)
  * Turn off the Ar laser using the key (if used)
  * Clean up immersion objectives
  * Close **ZEN**, shutdown the computer.
  * Turn off the metal halide fluorescence light source
  * Turn off the two switches: ''SYSTEMS/PC'' and ''COMPONENTS''
  * Wait 5 min. for Ar laser cooldown
  * Turn off the ''MAIN SWITCH''

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