zeiss_lsm_880
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zeiss_lsm_880 [2016/03/29 17:18] bioimaging created |
zeiss_lsm_880 [2020/02/27 15:35] bioimaging |
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| - | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_name.jpg"> | + | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_name_new.png"> |
| <table style="border:0px solid white;"> | <table style="border:0px solid white;"> | ||
| <tr style="border:0px solid white;"> | <tr style="border:0px solid white;"> | ||
| <td style="border:0px solid white;"> | <td style="border:0px solid white;"> | ||
| - | <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_wiki.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_wiki.jpg" width=300></a> | + | <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_wiki_new.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_880_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_lsm_880_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_lsm_880_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>: <a href="http://www.zeiss.com/microscopy/en_de/products/confocal-microscopes/lsm-880-with-airyscan-.html" target="_blank">LSM 880 with Airyscan</a><br> | + | <b>Model</b>: LSM 880 with Airyscan<br> |
| + | <b>Nickname</b>: "880"<br> | ||
| <b>Software</b>: ZEN 2.1 (black) <br> | <b>Software</b>: ZEN 2.1 (black) <br> | ||
| <b>Year</b>: 2016 <br> | <b>Year</b>: 2016 <br> | ||
| Line 14: | Line 15: | ||
|   <br> |   <br> | ||
|   <br> |   <br> | ||
| - | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/month.php?user=ZEISS5LIV&rid=12"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss LSM 880 Booking</a> | + |   <br> |
| + | → <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> | ||
| <br> | <br> | ||
| - | → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_7_live_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 7 Live 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> |
| - | <br> | + | |
| - | → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_7_live_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png"> Zeiss LSM 7 Live Usage Statistics</a> | + | |
| - |   </br> | + | |
| - | → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_7_live_repair"><img src="/facility/bioimaging/lib/exe/fetch.php?media=repair-icon.gif" width=16> Zeiss LSM 7 Live Repair History</a> | + | |
| <br> | <br> | ||
| - | → <a href="/facility/bioimaging/lib/exe/fetch.php?media=manual_lsm_5_live.pdf"><img src="/facility/bioimaging/lib/exe/fetch.php?media=adobe_pdf_icon.png"> Zeiss LSM 5 Live Operating Manual</a> | + | → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png"> Zeiss LSM 880 Usage Statistics</a> |
| - |   </p> | + | </p> |
| </td> | </td> | ||
| </tr> | </tr> | ||
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | The Zeiss LSM 7 Live //confocal line-scanning microscope// is a fast imaging system able to analyze high-speed processes with a time resolution of just a few microseconds. It is an upright microscope equipped with water dipping objectives specially suited for live samples in aqueous media. The scanning unit uses a laser beam with a rectangular cross-section to [[http://www.nature.com/nmeth/journal/v3/n11/thumbs/nmeth971-F2.jpg|illuminate a whole line in the sample]], instead of a single point. This makes image acquisition by its array CCD detectors faster than conventional PMT light detection but it also lowers the Z-resolution due to the usage of slits instead of conventional pinholes. Note that the system is equipped with two lasers (488 and 561 nm) but dual color imaging is not advised since there are no bandpass filters in the system (channel crosstalk is likely to occur). A UV laser controlled by galvanometric mirrors is available for precise ablation in user-defined regions of interest. The spatial resolution of this system 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]] but it is much faster than these two systems, achieving scanning speeds which may be even higher than the [[3i_marianas_sdc|3i Marianas SDC]] spinning disk if the acquisition is performed only in a few lines. 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.gif?|}} **If you are going to use the 561 nm laser please write '561' in the Event Field name** | + | {{: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. | ||
| + | |||
| + | ===== Booking Rules ===== | ||
| + | The Zeiss LSM 880 booking is divided in three daily slots: | ||
| + | | 9:00 to 13:00 | 13:00 to 17:00 | 17:00 to 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]]**. | ||
| + | - 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) | ||
| ===== System components ===== | ===== System components ===== | ||
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| ==== LASERs ==== | ==== LASERs ==== | ||
| ^ Laser Unit ^ Wavelength ^ Maximum Power ^ Current Status ^ | ^ Laser Unit ^ Wavelength ^ Maximum Power ^ Current Status ^ | ||
| - | | Sapphire 488-100 | 488 nm | 100 mW | ok | | + | | Diode 405-30 | 405 nm | 30 mW | ok | |
| - | | DPSS 561-40 | 561 nm | 40 mW | ok | | + | | Argon | 458, 488 and 514 nm | 25 mW | ok | |
| + | | DPSS 561-20 | 561 nm | 20 mW | ok | | ||
| + | | HeNe594 | 594 nm | 2 mW | ok | | ||
| + | | HeNe633 | 633 nm | 5 mW | ok | | ||
| ==== 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=440039-0000-000|W Achroplan Ph1]] | Water dipping | 0.30 | 3.1 | | + | | 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]] | |
| - | | 10x | [[https://www.micro-shop.zeiss.com/?s=2065696c9e13e&l=en&p=uk&f=o&a=v&m=s&id=440330-0000-000|Plan-Neofluar]] | Dry | 0.30 | 5.5 | | + | | 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 | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=421452-9900-000|W Plan-Apochromat]] | Water | 1.00 | 1.8 | | + | | 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 | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440090-9901-000|W Achroplan]] | Water dipping | 0.80 | 3.6 | | + | | 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]] | |
| - | | 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 | | + | |
| + | |||
| + | **Upon request:** | ||
| + | |||
| + | ^ 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/420852-9973-000/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]] | | ||
| + | | 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 ^ Emission ^ | ||
| - | | 2 | Green | [[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 | | + | | 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 | |
| - | | 3 | Red | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488015-0000-000&o=|FS15]] | 540-552 nm | > 590 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 | |
| - | | 4 | Blue (for ablation*) | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488049-9901-000&o=|FS49]] | none | 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 | 420-470 nm | |
| - | * Excitation filter has been removed | + | |
| - | ==== Filter and Dichroic sets ==== | + | ===== Microscope Turn On Procedure ===== |
| - | | {{zeiss5livefilters0.jpg?100}} | {{zeiss5livefilters1.jpg?100}} | | + | * Turn on the ''MAIN SWITCH'' |
| - | | Main configuration | Emission filters | | + | * Turn on the two switches: ''SYSTEMS/PC'' and ''COMPONENTS'' |
| + | {{zeiss_lsm_880_1.jpg?100}} | ||
| + | * Turn on the computer | ||
| + | * Login in to Windows (Bioimaging User) | ||
| + | * Check that the computer is connected to an ''Unidentified network'' (click on the network icon in the taskbar) | ||
| + | {{::network_icon.png?200}} | ||
| + | * Start the **ZEN** software | ||
| - | ===== Microscope Turn On Procedures ===== | + | ===== Microscope Turn Off Procedure ===== |
| - | + | If there is another user for this microscope in the __next hour__: | |
| - | ==== Normal Operation ==== | + | * Exit the ZEN software, leave the lasers on |
| - | + | * Log off the computer | |
| - | * The system must be turned on **30 min** before usage. | + | * Clean up immersion objectives |
| - | * To start it turn on the two main switches: ''SYSTEM/PC'' and ''COMPONENTS'' | + | |
| - | {{7live_switches.jpg?100|}} | + | |
| - | * Login to the computer, wait for the network icon to change to ''connected'' state | + | |
| - | * Start the **ZEN 2009** software. | + | |
| - | **After the 30 min:** | + | |
| - | * Turn on the metal halide lamp (white box to the left of the microscope). | + | |
| - | {{7live_excite.jpg?100|}} | + | |
| - | * You may now start using the system. | + | |
| - | + | ||
| - | ==== Live Imaging / Time Lapse Operation ==== | + | |
| - | + | ||
| - | * The system must be turned on **2 hours** before usage (in order to properly warm up scanning mirrors). | + | |
| - | * To start it turn on the two main switches: **SYSTEM/PC** and **COMPONENTS** | + | |
| - | * Login to the computer, wait for the network icon to change to ''connected'' state | + | |
| - | * Start the **ZEN 2009** software. | + | |
| - | **After the 2 hours:** | + | |
| - | * Turn on the metal halide lamp (white box to the left of the microscope). | + | |
| - | * You may now start using the system. | + | |
| - | + | ||
| - | ===== Microscope Turn Off procedures ===== | + | |
| - | If there's another user for this microscope in the next two hours: | + | |
| - | * Exit the **ZEN 2009** software, leave the lasers on and log off the computer. | + | |
| - | * Clean up immersion objectives. | + | |
| - | * **Make sure that there really is another user going to use the microscope.** | + | |
| Else: | Else: | ||
| - | * Turn off the lasers, wait 5 min for them to cooldown. | + | * Turn off the lasers on **ZEN** |
| - | * Exit the **ZEN 2009** software and shut down the computer. | + | * Clean up immersion objectives |
| - | * Clean up immersion objectives. | + | * Exit the ZEN software |
| - | * Turn off the metal halide lamp. | + | * Shutdown the computer |
| - | * Turn off the microscope. | + | * Turn off the two switches ''SYSTEMS/PC'' and ''COMPONENTS'' |
| - | * Turn off the two main switches: **SYSTEM/PC** and **COMPONENTS** | + | * **Wait 5 min. for Ar laser to cool down** |
| - | + | * Turn off the ''MAIN SWITCH'' | |
| - | ===== If you want to use the UV laser ablation system: ===== | + | |
| - | + | ||
| - | * Please contact the [[about|Bioimaging Facility]] (Ext: 47316). The ablation system requires specific training even for users that are already trained on the LSM 7 Live itself. | + | |
| ---- | ---- | ||
| - | |||
| [[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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