Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki_2022.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki_2022.png" width=300></a>
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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_map_new_p2a24.png">Room P2-A-24</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47219) <br> 
<b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">ZEISS Microscopy</a><br>
<b>Model</b>: Axiovert 200M<br>
<b>Nickname</b>: "Axiovert"<br>
<b>Software</b>: Metamorph 7.7.9.0<br>
<b>Year</b>: 2003 <br>
<b>SN</b>: ZCA522000494<br>
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Data will be deleted after: <b>1 month</b>
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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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===== Microscope overview =====

{{  :widefield_lightpath.png?0x300}}The Zeiss Axiovert 200M is a fully motorized inverted //widefield fluorescence microscope// ideal for live-cell imaging applications. 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 (Photometrics 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 [[https://huygens.i3s.up.pt/login.php|PPBI Huygens RM]] [[http://micro.magnet.fsu.edu/primer/digitalimaging/deconvolution/deconvolutionhome.html|deconvolution]] server 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_cell_observer|Zeiss Cell Observer]] with Definite Focus or the [[nikon_eclipse_ti|Nikon Eclipse Ti]]. If don't want to use deconvolution but need optical sectioning, check out one of the confocal systems such as the point scanners [[zeiss_lsm_710|Zeiss LSM 710]] and [[zeiss_lsm_880|Zeiss LSM 880]] or 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 [[colossus|Colossus]]. If you have FISH or other fixed fluorescence slides try the [[leica_dm5000b|Leica DM5000B]] instead.

  * **Microscope**: Zeiss Axiovert 200
  * **Monochrome Camera**: [[https://labs.pbrc.edu/cellbiology/documents/CoolSNAPHQ.pdf|Photometrics Coolsnap HQ CCD]]
  * **Color Camera**: [[https://www.leica-microsystems.com/products/microscope-cameras/p/leica-dfc450-c/|Leica DFC450]]

{{: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.

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

==== Filtersets ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Dichroic  ^  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  |  395 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  |  510 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  |  560 nm  |  575-640 nm  |
|  4  |  Far Red  |  [[http://www.semrock.com/SetDetails.aspx?id=2983|Cy5-4040C-000]]  |  608-648 nm  |  660 nm  |  672-712 nm  |
|  5  |  A  |  none  ||||

**Upon request:**

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Dichroic  ^  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  |  510 nm  |  > 515 nm  |

==== Objectives ====
^  Magnification  ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ^ (color)  10 μm =  ^  Reference  ^
|  10x  |  EC Plan-Neofluar  |  Air  |  0.30  |  5.20  |  1.00  |  10 pixels  |  23.3 pixels  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9900-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9900-000]]  |
|  20x  |  EC Plan-Neofluar  |  Air  |  0.50  |  2.00  |  0.48  |  21 pixels  |  46.7 pixels  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440340-9904-000/Objective-EC-Plan-Neofluar-20x-0.5|440340-9904-000]]  |
|  40x  |  EC Plan-NeoFluar  |  Air  |  0.75  |  0.71  |  0.24  |  41 pixels  |  93.7 pixels  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440350-9903-000/Objective-EC-Plan-Neofluar-40x-0.75|440350-9903-000]]  |
|  63x  |  Plan-Apochromat DIC  |  Oil  |  1.40  |  0.19  |  0.15  |  65 pixels  |  -  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420782-9900-000/Objective-Plan-Apochromat-63x-1.4-Oil-DIC-M27|420782-9900-000]]  |
|  100x  |  α Plan-Apochromat DIC  |  Oil  |  1.46  |  0.11  |  0.09  |  105 pixels  |  -  |  [[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]]  |


**Upon request:**
^  Magnification  ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ^  Reference  ^
|  1.25x  |  EC Epiplan-Neofluar  |  Air  |  0.035  |  4  |  7.58  |  1.3 pixels  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/422310-9900-000/Objective-EC-Epiplan-Neofluar-1.25x-0.03-M27|422310-9900-000]]  |
|  5x  |  EC Plan-NeoFluar  |  Air  |  0.16  |  18.5  |  2.00  |  5 pixels  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420330-9901-000/Objective-EC-Plan-Neofluar-5x-0.16-M27|420330-9901-000]]  |
|  40x  |  Plan-Neofluar Ph3  |  Oil  |  1.30  |  0.20  |  0.24  |  41 pixels  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440451-9903-000/Objective-EC-Plan-Neofluar-40x-1.30-Oil-Ph3|440451-9903-000]]  |
|  63x  |  Plan-Apochromat Ph3  |  Oil  |  1.4  |  0.19  |  0.15  |  65 pixels  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420781-9910-000/Objective-Plan-Apochromat-63-1.4-Oil-Ph3-M27|420781-9910-000]]  |

*** pixel sizes for binning = 1**

==== Cameras ====
^  Model  ^  Type  ^  Frame Size  ^  Pixel Size (µm)  ^
|  [[https://labs.pbrc.edu/cellbiology/documents/CoolSNAPHQ.pdf|Photometrics Coolsnap HQ]]  |  CCD  |  1392 x 1040  |  6.45 x 6.45  |
|  [[https://www.leica-microsystems.com/products/microscope-cameras/p/leica-dfc450-c/|Leica DFC450]]  |  color CCD  |  2560 × 1920  |  3.4 x 3.4  |

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

===== System Turn On Procedures =====
  * Turn on the fluorescent light power source (if used)
{{halide_710.jpg?80|}}
  * Turn on the Prior stage controller 
{{ax_prior_on.jpg?100|}}
  * Turn on the microscope
{{ax_mic_on.jpg?100|}}
  * Turn on the CCD camera
{{ax_cam_on.jpg?100|}}
  * Turn on the computer
  * Log in to Windows (Bioimaging User)
  * Start the Metamorph software
  * If the software doesn't detect the camera, power cycle the camera and try again
===== Microscope Turn Off procedures =====

If there is another user for this microscope in the __next hour__:
  * Leave the fluorescent lamp on (if used)
  * Clean up immersion objectives
Else:
  * Clean up immersion objectives
  * Shutdown the computer
  * Turn off the camera
  * Turn off the Prior stage controller
  * Turn off the microscope
  * Turn off the fluorescent lamp (if used)

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