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zeiss_axiovert_200m [2018/09/21 16:30]
bioimaging [Booking Rules]
zeiss_axiovert_200m [2020/03/01 18:05]
bioimaging [Filtersets]
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-<img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=axiovert_200m_name.jpg">+<img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_axiovert_200m_name_new.png">
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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>​+<a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_axiovert_200m_wiki_new.png"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_axiovert_200m_wiki_new.png" 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>​Location</​b>:​ <a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_axiovert_200m_map_new.png">​Room P2-A-22</​a>​ (<img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=phone_neg.png" width=15>​ 47217) <​br> ​
 <​b>​Manufacturer</​b>:​ <a href="​http://​www.zeiss.de/​micro"​ target="​_blank">​Carl Zeiss MicroImaging</​a><​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>​Model</​b>:​ Axiovert 200M<​br>​
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 <​b>​Year</​b>:​ 2003 <br> <​b>​Year</​b>:​ 2003 <br>
 <​b>​SN</​b>:​ ZCA522000494<​br>​ <​b>​SN</​b>:​ ZCA522000494<​br>​
 +&nbsp <br>
 &nbsp <br> &nbsp <br>
 &nbsp <br> &nbsp <br>
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  <​br>​  <​br>​
 &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>​ &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>​
- <​br>​ +</p>
-&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>​ +
- </p>+
 </td> </td>
 </tr> </tr>
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 ===== Microscope overview ===== ===== 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 (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 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_cell_observer|Zeiss Cell Observer]] with Definite Focus. If don't want to use deconvolution,​ 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 [[big_guy|Big Guy]] or [[colossus|Colossus]]. If you have FISH or other fixed fluorescence slides try the [[leica_dm5000b|Leica DM5000B]] instead.+{{  :​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 [[big_guy|Big Guy]] or [[colossus|Colossus]]. If you have FISH or other fixed fluorescence slides try the [[leica_dm5000b|Leica DM5000B]] instead.
  
   * **Microscope**:​ Zeiss Axiovert 200   * **Microscope**:​ Zeiss Axiovert 200
-  * **Camera**: Photometrics Coolsnap HQ CCD+  * **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.jpg?20|}} 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.+{{: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.
  
 ===== Booking Rules ===== ===== Booking Rules =====
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 ==== Filtersets ==== ==== Filtersets ====
  
-^  Position ​ ^  Filterset ​ ^  Reference ​ ^  Excitation ​ ^  Emission ​ ^ +^  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  |  > 397 nm  | +|  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  |  515-565 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  |  575-640 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  |  672-712 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  |  ​none  ​ ​none  ​|+|  5  |  A  |  none  ||||
  
 **Upon request:** **Upon request:**
  
-^  Position ​ ^  Filterset ​ ^  Reference ​ ^  Excitation ​ ^  Emission ​ ^ +^  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  |  > 515 nm  |+|  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 ==== ==== Objectives ====
-^  Magnification ​ ^  Model  ^  ​Type  ​^ ​ NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ^ (color ​camera)  10 μm =  ^ +^  Magnification ​ ^  Model  ^  ​Immersion ​ ​^ ​ NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ^ (color) ​ 10 μm =  ​^ ​ Reference ​ ^ 
-|  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. ​| ​ 1.00  |  10 pixels ​ |  23.3 pixels ​ | +|  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  ​| ​ [[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 ​ | +|  ​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  ​| ​ [[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 ​ | +|  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  ​| ​ [[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 ​ |  -  |+|  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]] ​ |
  
  
 **Upon request:** **Upon request:**
-^  Magnification ​ ^  Model  ^  ​Type  ​^ ​ NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ^ +^  Magnification ​ ^  Model  ^  ​Immersion ​ ​^ ​ NA  ^  WD (mm)  ^  pixel size* (μm)  ^  10 μm =  ​^ ​ Reference ​ ^ 
-|  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 ​ | +|  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  ​| ​ [[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 ​ | +|  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]]  ​
-|  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  ​|  ​ ​| ​ 0.48  ​|  ​21 pixels ​ +|  ​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]]  | 
-|  ​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  |  Plan-Apochromat Ph3  ​| ​ Oil  |  1. ​| ​ 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]]  | 
-|  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. ​| ​ 0.19  ​| ​ 0.15  ​|  ​65 pixels ​ |+|  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]] ​ |
  
 *** pixel sizes for binning = 1** *** 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 ==== ==== Stage ====
Line 87: Line 92:
  
 ===== System Turn On Procedures ===== ===== System Turn On Procedures =====
-  * Turn the fluorescence ​light power source ​on.+  * Turn on the fluorescent ​light power source
 {{halide_710.jpg?​80|}} {{halide_710.jpg?​80|}}
-  * Turn on the Uniblitz shutter controller (on the back). +  * Turn on the Prior stage controller ​
-{{ax_uniblitz1.jpg?​100|}}{{ax_uniblitz2.jpg?​100|}} +
-  * Turn the Prior microscope ​stage controller ​on. +
 {{ax_prior_on.jpg?​100|}} {{ax_prior_on.jpg?​100|}}
-  * Wait 10s then turn the microscope ​on.+  * Turn on the microscope
 {{ax_mic_on.jpg?​100|}} {{ax_mic_on.jpg?​100|}}
-  * Turn the camera ​on.+  * Turn on the CCD camera
 {{ax_cam_on.jpg?​100|}} {{ax_cam_on.jpg?​100|}}
-  * Turn the computer ​on. +  * Turn on the computer 
-  * Login to the computer ​(Axiovert 200M user). +  * Log in to Windows ​(Bioimaging User
-  * Start the Metamorph software. +  * Start the Metamorph software 
-  * If you need temperature control, turn on the control box +  * If the software doesn'​t detect the camerapower cycle the camera ​and try again
-{{heating2.jpg?​100|}} +
-  * If you need CO2open the valve and turn on CO2 supply on the control box +
-{{co2.jpg?​100|}} +
 ===== Microscope Turn Off procedures ===== ===== Microscope Turn Off procedures =====
-  * Turn off the computer. + 
-  * Turn off the camera. +If there is another user for this microscope in the __next hour__: 
-  * If you used any __immersion oil in the objectives, clean them up__. +  * Leave the fluorescent lamp on 
-  * Turn off the microscope stage controller. +  * Clean up immersion ​objectives 
-  * Turn off the shutter ​controller. +Else: 
-  * Turn off the microscope. +  * Clean up immersion objectives 
-  * If there'​s __another user__ for this microscope in the __next hour__ leave the mercury __lamp power source on, else turn it off__.  +  * Shutdown the computer 
-  * 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. +  * Turn off the camera 
 +  * Turn off the Prior stage controller 
 +  * Turn off the microscope 
 +  * Turn off the fluorescent ​lamp
  
 ---- ----
 [[resources|Back to the Resources page]] [[resources|Back to the Resources page]]
zeiss_axiovert_200m.txt · Last modified: 2020/07/21 23:27 by bioimaging