Bioimaging Wiki

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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axio_observer_map.jpg">Room P3-A-10</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47311) <br> 
<b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><br>
<b>Model</b>: Axio Observer<br>
<b>Software</b>: ZEN 2 (blue edition)<br>
<b>Year</b>: 2015 <br>
<b>SN</b>: 1026052056<br>
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&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/125"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss Axio Observer Booking</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_axio_observer_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss Axio Observer Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_axio_observer_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Zeiss Axio Observer Usage Statistics</a>
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===== Microscope overview =====
The Zeiss Axio Observer is a fully motorized inverted //widefield fluorescence microscope// ideal for live-cell imaging applications. It is equipped with a large cage incubator and a small stage incubator for temperature control and CO2 supply and with Definite Focus for hardware focus control during long time-lapse acquisitions. Using the ZEN 2 software you can control the motorized filters, shutters and stage to simultaneously set up multi-color time-lapse imaging experiments in multiple stage positions. Its sensitive cooled Axiocam 506m camera allows you to capture weak fluorescent signals and minimize photobleaching/photodamage in light sensitive samples. You can also perform Z-stack acquisition and use our [[http://micro.magnet.fsu.edu/primer/digitalimaging/deconvolution/deconvolutionhome.html|deconvolution]] server [[brainstorm|Brainstorm]] to significantly improve image resolution even in thick samples. If don't want to use deconvolution, check out one of the confocal systems such as the [[zeiss_lsm_710|Zeiss LSM 710]], the [[zeiss_lsm_880|Zeiss LSM 880]] or the [[3i_marianas_sdc|3i Marianas SDC]] spinning disk. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. If you have FISH or other fixed fluorescence slides try the [[leica_dm5000b|Leica DM5000B]] instead.

===== Booking Rules =====
The Zeiss Axio Observer 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 if they will use the incubator in the booking Description field (write down 37C INCUBATOR)
===== System components =====

==== Filtersets ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Emission  ^
|  1  |  HE GFP  |  [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=489038-9901-000&o=|FS38HE]]  |  450-490 nm  |  500-550 nm  |
|  2  |  HE DsRed  |  [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=489043-9901-000&o=|FS43HE]]  |  538-562 nm  |  570-640 nm  |
|  3  |  DAPI  |  [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=488049-9901-000&o=|FS49]]  |  335-383 nm  |  420-470 nm  |
|  4  |  Cy5  |  [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=488050-9901-000&o=|FS50]]  |  625-655 nm  |  665-715 nm  |
|  5  |  none  |  none  |  none  |  none  |
|  6  |  none  |  none  |  none  |  none  |

==== 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=2310860526ae9cb&l=en&p=uk&f=o&a=v&m=s&id=420650-9901-000|Plan-Apochromat]]  |  Dry  |  0.80  |  0.55  |
|  40x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440350-9903-000|EC Plan-NeoFluar]]  |  Dry  |  0.75  |  0.71  |
|  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  |

**Upon request:**
^  Magnification  ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
|  1.25x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=422310-9900-000|EC Epiplan-Neofluar]]  |  Dry  |  0.035  |  4  |
|  5x  |  [[https://www.micro-shop.zeiss.com/?s=571974962233f6&l=en&p=uk&f=o&a=v&m=s&id=420330-9901-000|EC Plan-NeoFluar]]  |  Dry  |  0.16  |  18.5  |
|  20x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440341-9904-000|EC Plan-Neofluar Ph 2]]  |  Dry  |  0.50  |  2  |
|  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  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440451-9903-000|Plan-Neofluar Ph3]]  |  Oil  |  1.30  |  0.20  |
|  63x  |  [[https://www.micro-shop.zeiss.com/?s=571858182d6d9d&l=en&p=uk&f=o&a=v&m=s&id=420781-9910-000|Plan-Apochromat Ph3]]  |  Oil  |  1.4  |  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  |  0.10  |  102 pixels  |

===== System Turn On Procedures =====
  * Turn on the HXP 120 fluorescence light.
{{zeiss_axio_observer_hxp.jpg?120|}}
  * Turn on the Temperature and CO2 controller (if you need them).
{{zeiss_axio_observer_temp.jpg?80|}}
  * If you need CO2, open the valve
{{zeiss_axio_observer_co2.jpg?80|}}
  * Turn on the Motorized Stage controller (A)
{{zeiss_axio_observer_a.jpg?height=150|}}
  * Turn on the Definite Focus controller (if you are going to use it)
{{zeiss_axio_observer_df.jpg?height=150|}}
  * Turn on the Microscope power supply (B)
{{zeiss_axio_observer_b.jpg?height=150|}}
  * Turn on the computer
  * Log in Windows with your lab user account (each lab has it's own area and login).
  * Start the **ZEN 2** software

===== Microscope Turn Off procedures =====
  * Exit the **ZEN 2** software
  * Turn off the computer
  * If you used any __immersion oil in the objectives, clean them up__.
  * Turn off the microscope power supply (B)
  * Turn off the Definite Focus controller (if used)
  * Turn off the Motorized Stage controller (A)
  * Turn off the Temperature and CO2 controller (if used)
  * If there's __another user__ for this microscope in the __next hour__ leave the mercury __lamp power source on, else turn it off__. 
  * Be extra careful in weekends and at the end of the day to make sure that the mercury lamp is not going to be turned on for an excessive time. 

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