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zeiss_cell_observer [2021/02/22 18:27] bioimaging |
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- | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_name.jpg"> | + | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_name_new.png"> |
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<a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_wiki_new.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_wiki_new.png" width=300></a> | <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_wiki_new.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_wiki_new.png" width=300></a> | ||
<td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
- | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_map.jpg">Room P3-A-10</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47331) <br> | + | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_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>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><br> | ||
<b>Model</b>: Axio Observer<br> | <b>Model</b>: Axio Observer<br> | ||
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  <br> |   <br> | ||
  <br> |   <br> | ||
- | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/125"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss Cell Observer Booking</a> | + | → <a href="https://my.agendo.science/calendar/?id=MjZjYWxlbmRhcnM="><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss Cell Observer Booking</a> |
<br> | <br> | ||
→ <a href="/facility/bioimaging/doku.php?id=zeiss_cell_observer_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss Cell Observer Quality Control</a> | → <a href="/facility/bioimaging/doku.php?id=zeiss_cell_observer_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss Cell Observer Quality Control</a> | ||
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===== Microscope overview ===== | ===== Microscope overview ===== | ||
- | The Zeiss Cell 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 point scanners [[zeiss_lsm_710|Zeiss LSM 710]] and [[zeiss_lsm_880|Zeiss LSM 880]] or 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 Cell Observer is a fully motorized inverted //widefield fluorescence microscope// ideal for live-cell imaging with fast acquisition speeds. It is equipped with a large cage incubator and a stage top incubator for temperature control and CO2 supply and with Definite Focus for hardware focus control during long time-lapse acquisitions. Using the ZEN software you can control the motorized filters, shutters and stage to simultaneously set up multi-color time-lapse imaging experiments in multiple stage positions. The stage is furthermore equipped with a piezo for fast Z displacement. You can choose between a HXP metal halide lamp or fast-switching LEDs for illumination. 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 [[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 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 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]]. |
* **Microscope**: [[http://www.zeiss.com/microscopy/en_de/products/light-microscopes/axio-observer-for-biology.html|Zeiss Axio Observer]] | * **Microscope**: [[http://www.zeiss.com/microscopy/en_de/products/light-microscopes/axio-observer-for-biology.html|Zeiss Axio Observer]] | ||
+ | * **LED Illumination**: [[https://www.zeiss.com/microscopy/int/products/microscope-components/lightsources/colibri-7.html?vaURL=www.zeiss.de/colibri|Zeiss Colibri 7]] | ||
* **Camera**: [[https://www.zeiss.com/microscopy/int/products/microscope-cameras/axiocam-506-mono.html|Axiocam 506 mono CCD]] | * **Camera**: [[https://www.zeiss.com/microscopy/int/products/microscope-cameras/axiocam-506-mono.html|Axiocam 506 mono CCD]] | ||
- | {{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 ===== | ||
- | The Zeiss Cell Observer booking is divided in three daily slots: | + | - **Users** can book at maximum **8 hours per week** |
- | | 9:00 to 13:00 | 13:00 to 17:00 | 17:00 to 21:00 | | + | - This usage restriction does not apply for **weekends** and for working days **before 9:00** and **after 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]]**. | - 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 ===== | ===== System components ===== | ||
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==== Filtersets ==== | ==== Filtersets ==== | ||
- | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Emission ^ | + | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ 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 | | + | | 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 | 495 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 | | + | | 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 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 | | + | | 3 | 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 | 660 nm | 665-715 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 | | + | | 4 | Analyzer DIC ||||| |
- | | 5 | none | none | none | none | | + | | 5 | QUAD | [[https://www.micro-shop.zeiss.com/en/us/shop/filterAssistant/filtersets/489090-9100-000|FS90HE]] | 370-400 nm | 405 nm | 410-440 nm | |
- | | 6 | none | none | none | none | | + | | ::: | ::: | ::: | 450-488 nm | 493 nm | 499-529 nm | |
+ | | ::: | ::: | ::: | 540-570 nm | 575 nm | 580-605 nm | | ||
+ | | ::: | ::: | ::: | 615-648 nm | 653 nm | 659-759 nm | | ||
+ | | 6 | 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 | 395 nm | 420-470 nm | | ||
==== Objectives ==== | ==== Objectives ==== | ||
- | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ | + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ 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 | 5.2 | | + | | 10x | EC Plan-Neofluar Ph1 | Air | 0.30 | 5.20 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420341-9911-000/Objective-EC-Plan-Neofluar-10x-0.30-Ph1-M27|420341-9911-000]] | |
- | | 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 | | + | | 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 | [[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 | | + | | 20x | EC Plan-Neofluar Ph 2 | Air | 0.50 | 2.00 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440341-9904-000/Objective-EC-Plan-Neofluar-20x-0.5-Ph2|440341-9904-000]] | |
- | | 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 | | + | | 40x | EC Plan-NeoFluar | Air | 0.75 | 0.71 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420360-9900-000/Objective-EC-Plan-Neofluar-40x-0.75-M27|420360-9900-000]] | |
+ | | 63x | Plan-Apochromat DIC | Oil | 1.40 | 0.19 | [[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]] | | ||
+ | | 63x | Plan-Apochromat Ph3 | Oil | 1.40 | 0.19 | [[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]] | | ||
**Upon request:** | **Upon request:** | ||
- | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ | + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ 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 | | + | | 1.25x | EC Epiplan-Neofluar | Air | 0.035 | 4.00 | [[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 | | + | | 5x | EC Plan-NeoFluar | Air | 0.16 | 18.50 | [[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 | 2 | | + | | 10x | EC Plan-Neofluar | Air | 0.30 | 5.20 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9901-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9901-000]] | |
- | | 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 | Plan-Apochromat Corr | Air | 0.95 | 0.25 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420660-9970-000/Objective-Plan-Apochromat-40x-0.95-Corr-M27|420660-9970-000]] | |
- | | 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 | | + | | 40x | Plan-Neofluar Ph3 | Oil | 1.30 | 0.20 | [[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 | [[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 | 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]] | |
- | | 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 | | + | |
+ | ==== Camera ==== | ||
+ | ^ Model ^ Type ^ Frame Size ^ Pixel Size (µm) ^ | ||
+ | | [[https://www.zeiss.com/microscopy/int/products/microscope-cameras/axiocam-506-mono.html|Axiocam 506 mono]] | CCD | 2752 x 2208 | 4.54 x 4.54 | | ||
===== System Turn On Procedures ===== | ===== System Turn On Procedures ===== | ||
- | * Turn on the HXP 120 fluorescence light. | + | |
+ | * Select LED or HXP light source on the manual path deflecting mirror (in the back of the microscope) | ||
+ | {{:cell_observer_led_selected.png?150|}} {{:cell_observer_hxp_selected.png?150|}} | ||
+ | * Turn on the HXP 120 fluorescent light source (if used) | ||
{{zeiss_axio_observer_hxp.jpg?120|}} | {{zeiss_axio_observer_hxp.jpg?120|}} | ||
- | * Turn on the Temperature and CO2 controller (if you need them). | + | * Turn on the temperature and CO2 controller (if you need them). |
{{zeiss_axio_observer_temp.jpg?80|}} | {{zeiss_axio_observer_temp.jpg?80|}} | ||
- | * If you need CO2, open the valve | + | * If you need CO2, open the Cell Observer CO2 valve (the main valve should always be open) |
- | {{zeiss_axio_observer_co2.jpg?80|}} | + | {{::p3a10_co2_valves.png?200|}} |
- | * Turn on the Motorized Stage controller (A) | + | * Turn on the motorized stage controller (**A** on remote control) |
{{zeiss_axio_observer_a.jpg?height=150|}} | {{zeiss_axio_observer_a.jpg?height=150|}} | ||
* Turn on the Definite Focus controller (if you are going to use it) | * Turn on the Definite Focus controller (if you are going to use it) | ||
{{zeiss_axio_observer_df.jpg?height=150|}} | {{zeiss_axio_observer_df.jpg?height=150|}} | ||
- | * Turn on the Microscope power supply (B) | + | * Turn on the microscope power supply (**B** on remote control) |
{{zeiss_axio_observer_b.jpg?height=150|}} | {{zeiss_axio_observer_b.jpg?height=150|}} | ||
* Turn on the computer | * Turn on the computer | ||
- | * Log in Windows with Cell Observer user account. | + | * Log in to Windows (Biomaging User). |
- | * Start the **ZEN 2** software | + | * Start the **ZEN** software |
+ | * Select HXP or LED Workspace in ZEN (top left) | ||
+ | {{::workspace_selection.png?250|}} | ||
+ | |||
+ | **Warning**: If you need to change the stage adapter, please contact the <html><a href="/facility/bioimaging/doku.php?id=about">Bioimaging Unit</a> (<b>imm-bioimaging@medicina.ulisboa.pt</b> | <img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47222</html>) | ||
===== Microscope Turn Off procedures ===== | ===== Microscope Turn Off procedures ===== | ||
- | * Exit the **ZEN 2** software | + | If there is another user for this microscope is the __next hour__: |
+ | * Leave the fluorescent light source on | ||
+ | * Clean up immersion objectives | ||
+ | Else: | ||
+ | * Exit the ZEN software | ||
* Turn off the computer (not the UPS!) | * Turn off the computer (not the UPS!) | ||
{{:cell_observer_pc_vs_ups.png?250|}} | {{:cell_observer_pc_vs_ups.png?250|}} | ||
- | * If you used any __immersion oil in the objectives, clean them up__. | + | * Clean up immersion objectives |
- | * Turn off the microscope power supply (B) | + | * Turn off the microscope power supply (**B** on remote control) |
* Turn off the Definite Focus controller (if used) | * Turn off the Definite Focus controller (if used) | ||
- | * Turn off the Motorized Stage controller (A) | + | * Turn off the motorized stage controller (**A** on remote control) |
* Turn off the Temperature and CO2 controller (if used) | * 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__. | + | * Close the Cell Observer CO2 valve (if used) |
- | * 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 HXP 120 fluorescent light source (if used) |
---- | ---- | ||
- | [[resources|Back to the Resources page]] | + | [[resources|Back to the Equipment page]] |