zeiss_lsm_880
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zeiss_lsm_880 [2020/01/14 18:06] bioimaging [Microscope Turn On Procedure] |
zeiss_lsm_880 [2020/06/01 19:22] bioimaging |
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| - | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/128"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss LSM 880 Booking</a> | + | → <a href="https://my.agendo.science/calendar/?id=OGNhbGVuZGFycw=="><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss LSM 880 Booking</a> |
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| → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 880 Quality Control</a> | → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 880 Quality Control</a> | ||
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | {{ :airyscan.jpg?0x380}} The Zeiss LSM 880 with Airyscan is a //confocal point-scanning microscope// able to generate high-resolution three-dimensional images of thick specimens with high sensitivity and low photodamage. It is an inverted microscope especially suitable for live cell imaging and photobleaching experiments, equipped with a large size incubator for temperature control and CO2 supply and with Definite Focus for hardware focus control during long time-lapse acquisitions. Its [[http://www.zeiss.com/content/dam/Microscopy/Products/laser-scan/LSM%20880/zeiss-lsm880-airyscan-principle-672.jpg|Airyscan]] 32 channel area detector is able to collect all light from an Airy pattern simultaneously, allowing for increased resolution (140 nm laterally and 400 nm axially, at 488 nm). Its scanning unit further includes two PMT detectors (one of them cooled) as well as a GaAsP detector for increased sensitivity (45% QE compared to ~25% QE for conventional PMT). It is equipped with lasers from violet to far red (405, 458, 488, 514, 561, 594 and 633 nm excitation wavelengths). 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]] or the [[zeiss_axiovert_200m|Zeiss Axiovert 200M]]. Image resolution with Airyscan is higher than the [[zeiss_lsm_710|Zeiss LSM 710]] and detection sensitivity is also higher than faster confocal systems such as 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]]. | + | {{ :airyscan_beampath.png?400}} The Zeiss LSM 880 with Airyscan is a //confocal point-scanning microscope// able to generate high-resolution three-dimensional images of thick specimens with high sensitivity, high speed and low photodamage. It is an inverted microscope especially suitable for live cell imaging and photobleaching experiments, equipped with a large size incubator for temperature control and CO2 supply and with Definite Focus for hardware focus control during long time-lapse acquisitions. The stage is motorized and furthermore equipped with a piezo for Z displacement so fast 4D imaging is possible in multiple stage positions. Its [[http://www.zeiss.com/content/dam/Microscopy/Products/laser-scan/LSM%20880/zeiss-lsm880-airyscan-principle-672.jpg|Airyscan]] 32 channel area detector is able to collect all light from an Airy pattern simultaneously, allowing for increased resolution (140 nm laterally and 400 nm axially, at 488 nm). It can also be used in [[https://www.youtube.com/watch?v=ln6VUmLzLLc|Fast Mode]], which enables parallel excitation and detection of four image pixels. The result is a speed improvement by a factor of four, while maintaining the sensitivity of Airyscan and 1.5x resolution improvement. Its scanning unit further includes two PMT detectors (one of them cooled) as well as a GaAsP detector for increased sensitivity (45% QE compared to ~25% QE for conventional PMT). It is equipped with lasers from violet to far red (405, 458, 488, 514, 561, 594 and 633 nm excitation wavelengths). |
| + | {{::fast_airyscan.png?420 }}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]] or the [[zeiss_axiovert_200m|Zeiss Axiovert 200M]]. Image resolution with Airyscan is higher than the [[zeiss_lsm_710|Zeiss LSM 710]] and detection sensitivity is also higher than 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]]. | ||
| - | {{:info.png?25|}} Click on the image on the right to see the system beam path in higher detail | + | {{:info.png?25|}} Click [[https://imm.medicina.ulisboa.pt/facility/bioimaging/lib/exe/fetch.php?cache=&media=airyscan.jpg|here]] to see the system beam path in higher detail |
| {{: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. | {{: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. | ||
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| - Users must indicate which lasers they are going to use in the booking Description field | - Users must indicate which lasers they are going to use in the booking Description field | ||
| - Users must indicate if they will use the incubator in the booking Description field (write down 37C INCUBATOR) | - Users must indicate if they will use the incubator in the booking Description field (write down 37C INCUBATOR) | ||
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| ===== System components ===== | ===== System components ===== | ||
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| ==== 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 | 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]] | |
| | 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 | C-Apochromat Corr | Water | 1.20 | 0.28 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/421767-9970-000/Objective-C-Apochromat-40x-1.20-W-Corr-M27|421767-9970-000]] | | | 40x | C-Apochromat Corr | Water | 1.20 | 0.28 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/421767-9970-000/Objective-C-Apochromat-40x-1.20-W-Corr-M27|421767-9970-000]] | | ||
| - | | 63x | Plan-Apochromat DIC | Oil | 1.40 | 0.19 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440762-9904-000/Objective-Plan-Apochromat-63x-1.40-Oil-DIC|440762-9904-000]] | | + | | 63x | Plan-Apochromat DIC | Oil | 1.40 | 0.19 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420782-9900-799/Objective-Plan-Apochromat-63x-1.4-Oil-DIC-M27|420782-9900-799]] | |
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| ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
| - | | 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]] | | + | | 25x | LCI Plan-Neofluar Corr DIC | Oil/Glyc/W | 0.80 | 0.21 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420852-9973-000/Objective-LCI-Plan-Neofluar-25x-0.8-Imm-Corr-DIC-M27|420852-9972-000]] | |
| | 40x | Plan-Apochromat Corr | Dry | 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 | Dry | 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]] | | ||
| | 100x | α Plan-Apochromat DIC | Oil | 1.46 | 0.11 | [[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]] | | | 100x | α Plan-Apochromat DIC | Oil | 1.46 | 0.11 | [[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]] | | ||
| ==== Filtersets (Ocular) ==== | ==== Filtersets (Ocular) ==== | ||
| - | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Emission ^ | + | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ |
| - | | 1 | Green | [[https://www.micro-shop.zeiss.com/?s=7139069fdc79c&l=en&p=us&f=f&a=v&b=f&id=489038-9901-000&o=|FS38HE]] | 450-490 nm | 500-550 nm | | + | | 1 | Green | [[https://www.micro-shop.zeiss.com/?s=7139069fdc79c&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 | Red | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=000000-1114-101&o=|FS43]] | 533-558 nm | 570-640 nm | | + | | 2 | Red | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=000000-1114-101&o=|FS43]] | 533-558 nm | 570 nm | 570-640 nm | |
| - | | 3 | Blue | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488049-9901-000&o=|FS49]] | G 365 nm | 420-470 nm | | + | | 3 | Blue | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488049-9901-000&o=|FS49]] | G 365 nm | 395 nm | 420-470 nm | |
| ===== Microscope Turn On Procedure ===== | ===== Microscope Turn On Procedure ===== | ||
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| * Turn on the computer | * Turn on the computer | ||
| * Login in to Windows (Bioimaging User) | * Login in to Windows (Bioimaging User) | ||
| - | * Check that the computer is connected to an ''Unidentified network'' (click on the network icon in the taskbar) | + | * Start the **ZEN Black** software |
| - | {{::network_icon.png?200}} | + | |
| - | * Start the **ZEN** software | + | |
| ===== Microscope Turn Off Procedure ===== | ===== Microscope Turn Off Procedure ===== | ||
| - | If there's another user for this microscope in the next hour: | + | If there is another user for this microscope in the __next hour__: |
| - | * Close the software, leave the lasers on and log off the computer. | + | * Exit the ZEN software, leave the lasers on |
| - | * Clean up any objective where you used immersion oil. | + | * Log off the computer |
| - | * **Make sure that there really is another user going to use the microscope.** | + | * Clean up immersion objectives |
| Else: | Else: | ||
| * Turn off the lasers on **ZEN** | * Turn off the lasers on **ZEN** | ||
| - | * Clean up any objective were you used immersion oil. | + | * Clean up immersion objectives |
| - | * Exit the software, shut down the computer. | + | * Exit the ZEN software |
| - | * Turn off the main switches ''SYSTEMS/PC'' and ''COMPONENTS'' | + | * Shutdown the computer |
| - | * Wait 5 min. for Ar laser cooldown | + | * Turn off the two switches ''SYSTEMS/PC'' and ''COMPONENTS'' |
| - | * Turn off the ''MAIN SWITCH''. | + | * **Wait 5 min. for Ar laser to cool down** |
| + | * Turn off the ''MAIN SWITCH'' | ||
| ---- | ---- | ||
| [[resources|Back to the Resources page]] | [[resources|Back to the Resources page]] | ||
zeiss_lsm_880.txt · Last modified: 2024/01/28 11:47 by bioimaging
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