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zeiss_cell_observer [2020/01/01 22:09] bioimaging [System Turn On Procedures] |
zeiss_cell_observer [2020/10/27 18:23] bioimaging |
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- | → <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> |
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→ <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 [[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]]. 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 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 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 [[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]] | ||
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===== 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 | 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 | |
- | | 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 | 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 | |
- | | 5 | none | none | none | none | | + | | 5 | Empty ||||| |
- | | 6 | none | none | none | none | | + | | 6 | Empty ||||| |
==== Objectives ==== | ==== Objectives ==== | ||
^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
- | | 10x | EC Plan-Neofluar | Air | 0.30 | 5.2 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9901-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9901-000]] | | + | | 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 | 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]] | | ||
| 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]] | | | 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 | EC Plan-NeoFluar | Air | 0.75 | 0.71 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440350-9903-000/Objective-EC-Plan-Neofluar-40x-0.75|440350-9903-000]] | | + | | 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 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 ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
- | | 1.25x | EC Epiplan-Neofluar | Air | 0.035 | 4 | [[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]] | | + | | 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 | EC Plan-NeoFluar | Air | 0.16 | 18.5 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420330-9901-000/Objective-EC-Plan-Neofluar-5x-0.16-M27|420330-9901-000]] | | + | | 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 | EC Plan-Neofluar Ph 2 | Air | 0.50 | 2 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440341-9904-000/Objective-EC-Plan-Neofluar-20x-0.5-Ph2|440341-9904-000]] | | + | | 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 | 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 | 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 | 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]] | | | 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 | Plan-Apochromat Ph3 | Oil | 1.4 | 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]] | | ||
| 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 | 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]] | | ||
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| [[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 | | | [[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. | + | * Turn on the HXP 120 fluorescent light source |
{{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 valve | ||
{{zeiss_axio_observer_co2.jpg?80|}} | {{zeiss_axio_observer_co2.jpg?80|}} | ||
- | * 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 (Biomaging User). | + | * Log in to Windows (Biomaging User). |
- | * Start the **ZEN 2** software | + | * Start the **ZEN** software |
+ | |||
+ | **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__. | + | * Turn off the HXP 120 fluorescent light source |
- | * 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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[[resources|Back to the Resources page]] | [[resources|Back to the Resources page]] |