Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki.jpg" width=300></a>
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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_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>: Axiovert 200M<br>
<b>Software</b>: Metamorph 7.7.9.0<br>
<b>Year</b>: 2003 <br>
<b>SN</b>: ZCA522000494<br>
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&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/3"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss Axiovert 200M Booking</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_axiovert200m_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss Axiovert 200M Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_axiovert_200m_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Zeiss Axiovert 200M Usage Statistics</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_axiovert_200m_repair"><img src="/facility/bioimaging/lib/exe/fetch.php?media=repair-icon.gif" width=16> Zeiss Axiovert 200M Repair History</a>
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===== Microscope overview =====

The Zeiss Axiovert 200M is a fully motorized inverted //widefield fluorescence microscope// ideal for live-cell imaging applications. It is equipped with a small stage incubator for temperature control and CO2 supply. Using the Metamorph 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 CCD camera (Roper Scientific Coolsnap HQ CCD) 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 you are performing long time-lapse experiments in live samples (aqueous media) and need hardware focus control, check the [[zeiss_axio_observer|Zeiss Axio Observer]] with Definite Focus. If don't want to use deconvolution, check out one of the confocal systems such as the [[zeiss_lsm_710|Zeiss LSM 710]], [[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 Axiovert 200M 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 =====

==== Filtersets ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Emission  ^
|  1  |  Blue  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488001-9901-000&o=|FS01]]  |  359-371 nm  |  > 397 nm  |
|  2  |  Green BP  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488010-9901-000&o=|FS10]]  |  450-490 nm  |  515-565 nm  |
|  3  |  Red  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488020-9901-000&o=|FS20]]  |  540-552 nm  |  575-640 nm  |
|  4  |  Far Red  |  [[http://www.semrock.com/SetDetails.aspx?id=2983|Cy5-4040C-000]]  |  608-648 nm  |  672-712 nm  |
|  5  |  A  |  none  |  none  |  none  |

**Upon request:**

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Emission  ^
|  2  |  Green LP  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488009-9901-000&o=|FS09]]  |  450-490 nm  |  > 515 nm  |

==== Objectives ====
^  Magnification  ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ^ (color camera)  10 μm =  ^
|  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  |  1.00  |  10 pixels  |  23.3 pixels  |
|  20x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420650-9901-000|EC Plan-Neofluar]]  |  Dry  |  0.50  |  2  |  0.48  |  21 pixels  |  46.7 pixels  |
|  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  |  0.24  |  41 pixels  |  93.7 pixels  |
|  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  |  0.15  |  65 pixels  |  -  |


**Upon request:**
^  Magnification  ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ^
|  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  |  7.58  |  1.3 pixels  |
|  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  |  2.00  |  5 pixels  |
|  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  |  0.48  |  21 pixels  |
|  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  |  0.24  |  41 pixels  |
|  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  |  0.15  |  65 pixels  |
|  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  |

*** pixel sizes for binning = 1**

==== Stage ====
For automated acquisition of 96 well plates use:
X = -9014 μm
Y = -8995 μm

===== System Turn On Procedures =====
  * Turn the fluorescence light power source on.
{{halide_710.jpg?80|}}
  * Turn on the Uniblitz shutter controller (on the back).
{{ax_uniblitz1.jpg?100|}}{{ax_uniblitz2.jpg?100|}}
  * Turn the Prior microscope stage controller on. 
{{ax_prior_on.jpg?100|}}
  * Wait 10s then turn the microscope on.
{{ax_mic_on.jpg?100|}}
  * Turn the camera on.
{{ax_cam_on.jpg?100|}}
  * Turn the computer on.
  * Login to the computer (each group has it's own area and login).
  * Start the Metamorph software.
  * If you need temperature control, turn on the control box
{{heating2.jpg?100|}}
  * If you need CO2, open the valve and turn on CO2 supply on the control box
{{co2.jpg?100|}}

===== Microscope Turn Off procedures =====
  * Turn off the computer.
  * Turn off the camera.
  * If you used any __immersion oil in the objectives, clean them up__.
  * Turn off the microscope stage controller.
  * Turn off the shutter controller.
  * Turn off the microscope.
  * 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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