3i_marianas_sdc
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3i_marianas_sdc [2019/12/31 18:17] bioimaging |
3i_marianas_sdc [2020/01/21 18:27] bioimaging [Microscope Turn Off Procedure] |
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| - | {{ :spinning_disk.jpg?0x300|}} The 3i Marianas SDC //spinning disk confocal microscope// is a fast imaging system which provides a trade-off between confocality, resolution and speed. It is an inverted microscope ideal for live cell applications which require fast acquisition speeds rather than high resolution images. The scanning unit achieves confocality by directing light through a spinning disk with many small pinholes. Images are then acquired with a sensitive EMCCD which allows for very small exposure times but is limited in resolution to 512x512 pixels. The stage is motorized and furthermore equipped with a piezo for Z displacement so fast 4D imaging is possible in multiple stage positions. The system is also equipped with a large size incubator for temperature control and a small stage incubator for CO2 supply. With this system you can perform optical sectioning of fluorescent samples which are too thick for a widefield system such as the [[zeiss_cell_observer|Zeiss Cell Observer]]. Even though its resolution is not as high as a //point-scanning confocal system// such as the [[zeiss_lsm_880|Zeiss LSM 880]] or [[zeiss_lsm_710|Zeiss LSM 710]], it is much faster than these two systems. You can also check the [[zeiss_cell_observer_sd|Zeiss Cell Observer SD]] if you need a spinning disk with a Definite Focus system for long time-lapse acquisitions. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. | + | {{ :spinning_disk.png|}} The 3i Marianas SDC //spinning disk confocal microscope// is a fast imaging system which provides a trade-off between confocality, resolution and speed. It is an inverted microscope ideal for live cell applications which require fast acquisition speeds rather than high resolution images. The scanning unit achieves confocality by directing light through a spinning disk with many small pinholes. Images are then acquired with a sensitive EMCCD which allows for very small exposure times but is limited in resolution to 512x512 pixels. The stage is motorized and furthermore equipped with a piezo for Z displacement so fast 4D imaging is possible in multiple stage positions. The system is also equipped with a large size incubator for temperature control and a small stage incubator for CO2 supply. With this system you can perform optical sectioning of fluorescent samples which are too thick for a widefield system such as the [[zeiss_cell_observer|Zeiss Cell Observer]]. Even though its resolution is not as high as a //point-scanning confocal system// such as the [[zeiss_lsm_880|Zeiss LSM 880]] or [[zeiss_lsm_710|Zeiss LSM 710]], it is much faster than these two systems. You can also check the [[zeiss_cell_observer_sd|Zeiss Cell Observer SD]] if you need a spinning disk with a Definite Focus system for long time-lapse acquisitions. 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]] | ||
| - | * **Confocal scanner**: [[http://www.andor.com/microscopy_systems/peripherals/confocal_scanners/|Yokogawa CSU-x1]] | + | * **Confocal scanner**: [[https://www.yokogawa.com/eu/solutions/products-platforms/life-science/spinning-disk-confocal/csu-x1-confocal-scanner-unit/|Yokogawa CSU-X1]] |
| - | * **Camera**: [[http://www.photometrics.com/products/emccdcams/evolve/512.php|Evolve 512 EMCCD]] | + | * **Camera**: [[https://www.photometrics.com/wp-content/uploads/2019/10/Evolve512-Datasheet.pdf|Photometrics Evolve 512 EMCCD]] |
| - | {{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. |
| ===== System components ===== | ===== System components ===== | ||
| Line 47: | Line 47: | ||
| ==== Objectives ==== | ==== Objectives ==== | ||
| - | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ | + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ |
| - | | 20x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420650-9901-000|Plan-Apochromat]] | Dry | 0.8 | 0.55 | | + | | 20x | Plan-Apochromat | Air | 0.8 | 0.55 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420650-9902-000/Objective-Plan-Apochromat-20x-0.8-M27#|420650-9902-000]] | |
| - | | 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 | | + | | 63x | Plan-Apochromat | Oil | 1.40 | 0.19 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420780-9900-000/Objective-Plan-Apochromat-63x-1.4-Oil-M27|420780-9900-000]] | |
| - | | 100x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420790-9901-000|Plan-Apochromat]] | Oil | 1.40 | 0.17 | | + | | 100x | Plan-Apochromat | Oil | 1.40 | 0.17 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420790-9901-000/Objective-Plan-Apochromat-100x-1.40-Oil-M27|420790-9901-000]] | |
| **Upon request:** | **Upon request:** | ||
| - | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ | + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ |
| - | | 25x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420852-9972-000|LCI Plan-NeoFluar]] | Oil/Glyc/W | 0.8 | 0.21 | | + | | 25x | LCI Plan-Neofluar Corr DIC | Oil/Glyc/W | 0.80 | 0.21 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420852-9972-000/Objective-LCI-Plan-Neofluar-25x-0.8-Imm-Corr-DIC-M27|420852-9972-000]] | |
| - | | 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]] | |
| - | ==== Emission Filtersets ==== | + | |
| + | ==== Emission Filters ==== | ||
| ^ Setting ^ Transmission ^ | ^ Setting ^ Transmission ^ | ||
| | 1: 445 | 422-477 nm | | | 1: 445 | 422-477 nm | | ||
| Line 69: | Line 70: | ||
| | 10: 445 | 422-477 nm | | | 10: 445 | 422-477 nm | | ||
| + | ==== Camera ==== | ||
| + | ^ Model ^ Frame Size ^ Pixel Size (µm) ^ Quantum Efficiency ^ | ||
| + | | [[https://www.photometrics.com/wp-content/uploads/2019/10/Evolve512-Datasheet.pdf|Photometrics Evolve 512 EMCCD]] | 512 x 512 | 16 x 16 | > 90% | | ||
| ===== Microscope Turn On Procedure ===== | ===== Microscope Turn On Procedure ===== | ||
| * Check that the main power supply switch is on (should be on by default) | * Check that the main power supply switch is on (should be on by default) | ||
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| * Turn on the fluorescence lamp (if needed) | * Turn on the fluorescence lamp (if needed) | ||
| {{marianas_power5.jpg?|}} | {{marianas_power5.jpg?|}} | ||
| - | * Turn on the power switch distributor (green) and the component power switches (red) **one at a time** | + | * Turn on the power switch distributor (green) and the component power switches (red) **one at a time** from left to right |
| {{marianas_power3.jpg?|}} | {{marianas_power3.jpg?|}} | ||
| - | * Turn on the secondary power supply switch | + | * Turn on the secondary power supply switch (white socket outlet underneath the touchscreen) |
| - | {{marianas_power2.jpg?|}} | + | |
| * Turn on the Yokogawa spinning disk scanner (using the key) | * Turn on the Yokogawa spinning disk scanner (using the key) | ||
| {{marianas_power4.jpg?|}} | {{marianas_power4.jpg?|}} | ||
| Line 88: | Line 91: | ||
| * Start the **SlideBook** software | * Start the **SlideBook** software | ||
| - | **Warning**: If you need to change the stage adaptor, please contact the <html><a href="/facility/bioimaging/doku.php?id=about">Bioimaging Facility</a> (<b>imm-bioimaging@medicina.ulisboa.pt</b> | <img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47316</html>) | + | **Warning**: If you need to change the stage adaptor, 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 Procedure ===== | ===== Microscope Turn Off Procedure ===== | ||
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| * Exit the **SlideBook** software | * Exit the **SlideBook** software | ||
| * Switch off the computer | * Switch off the computer | ||
| - | * Turn off all system components (follow the Turn On Procedure in reverse) | + | * Turn off incubation in the touchscreen (if used) |
| + | * Turn off CO2 (if used) | ||
| + | * Switch all lasers off (first on the switch and finally by turning the key) | ||
| + | * Turn off the fluorescent lamp | ||
| + | * Turn off the secondary power supply switch (white socket outlet underneath the touchscreen) | ||
| + | * Turn off the component power switches (red) **one at a time** from right to left and the power switch distributor (green) | ||
| ---- | ---- | ||
| [[resources|Back to the Resources page]] | [[resources|Back to the Resources page]] | ||
3i_marianas_sdc.txt · Last modified: 2024/04/01 23:50 by bioimaging
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