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zeiss_lsm_710 [2016/04/05 01:48] bioimaging created |
zeiss_lsm_710 [2024/01/18 01:18] (current) bioimaging |
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- | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_710_name.jpg"> | + | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_710_name_new.png"> |
- | <table border="0" align="top"> | + | <table style="border:0px solid white;"> |
- | <tr> | + | <tr style="border:0px solid white;"> |
- | <td> | + | <td style="border:0px solid white;"> |
- | <a href="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/zeiss_lsm_710_wiki.jpg"><img src="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/zeiss_lsm_710_wiki.jpg" width=300></a> | + | <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_710_wiki_2022.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_710_wiki_2022.png" width=300></a> |
- | <td>   </td> | + | <td style="border:0px solid white;"> <p style="line-height:1.8"> |
- | <td> <b>Location</b>: <a href="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/zeiss_lsm_710_map.jpg">Room P3-A-10</a> (<img src="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/telephone_icon.gif" width=15> 47311) <p> | + | <b>Location</b>: no longer available <br> |
- | <b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><p> | + | <b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">ZEISS Microscopy</a><br> |
- | <b>Model</b>: <a href="http://microscopy.zeiss.com/microscopy/en_de/products/confocal-microscopes/confocal-microscopy-lsm-710.html" target="_blank">LSM 710</a><p> | + | <b>Model</b>: LSM 710<br> |
- | <b>Software</b>: ZEN 2009<p> | + | <b>Nickname</b>: "710"<br> |
- | <b>Year</b>: 2010 <p> | + | <b>Software</b>: ZEN 2012 SP5 FP3 (black)<br> |
- | <b>SN</b>: 2503000400 <p> | + | <b>Year</b>: 2010 <br> |
- |   <p> | + | <b>SN</b>: 2503000400 <br> |
- | → <a href="http://www.imm.fm.ul.pt/booking/month.php?user=ZEISSLSM7&rid=51"><img src="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/date.png"> Zeiss LSM 710 Booking</a> | + |   <br> |
- | <p> | + | Data will be deleted after: <b>3 months</b> |
- | → <a href="http://www.imm.fm.ul.pt/wiki/doku.php?id=bioimaging:zeiss_710_quality"><img src="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/list-icon.png" width=18> Zeiss LSM 710 Quality Control</a> | + |   <br> |
- | <p> | + |   <br> |
- | → <a href="http://www.imm.fm.ul.pt/wiki/doku.php?id=bioimaging:710usage"><img src="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/chart_line.png"> Zeiss LSM 710 Usage Report</a> | + |   <br> |
- | <p> | + | → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_710_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 710 Quality Control</a> |
- | → <a href="http://www.imm.fm.ul.pt/wiki/doku.php?id=bioimaging:710repair"><img src="http://www.imm.fm.ul.pt/wiki/data/media/bioimaging/repair-icon.gif" width=16> Zeiss LSM 710 Repair History</a> | + | <br> |
- |   <p> | + | → <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_710_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png"> Zeiss LSM 710 Usage Statistics</a> |
- | <p> | + | </p> |
- | → <a href="http://www.imm.fm.ul.pt/wiki/lib/exe/fetch.php?media=bioimaging:manual_lsm_710.pdf"><img src="http://www.imm.fm.ul.pt/wiki/data/media/groups/bioimage/adobe_pdf_icon.png" width=16> Zeiss LSM 710 Operating Manual</a> | + | |
- |   <p> | + | |
- | <p> | + | |
- | → <a href="http://www.imm.fm.ul.pt/wiki/lib/exe/fetch.php?media=bioimaging:manual_lsm_710_quick_guide.pdf"><img src="http://www.imm.fm.ul.pt/wiki/data/media/groups/bioimage/adobe_pdf_icon.png" width=16> Zeiss LSM 710 Quick Guide</a> | + | |
- |   <p> | + | |
</td> | </td> | ||
</tr> | </tr> | ||
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===== Microscope overview ===== | ===== Microscope overview ===== | ||
- | The Zeiss LSM 710 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 specially suitable for live cell imaging and photobleaching experiments, equipped with a large size incubator for temperature control (but no stage incubator for CO2 supply). Its scanning unit has a spectral recycling loop, two sensitive PMT detectors and a filter-free spectral separation module that can be continuously set over the entire wavelength range (i.e. you can specify which wavelength range you want to detect). It is equipped with lasers from violet to far red (405, 458, 488, 561 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 [[bioimaging:zeiss_axiovert_200m|Zeiss Axiovert 200M]]. Image resolution and detection sensitivity are higher than faster confocal systems such as a spinning disk or the [[bioimaging:zeiss_7_live|Zeiss LSM 7 Live]], but image acquisition is slower. If you need to use a 594 nm laser or a 40x water objective for higher working distances, check the [[bioimaging:zeiss_lsm510_meta|Zeiss LSM 510 META]] confocal point-scanning system. If your personal computer cannot handle all the data you collected, check out the [[bioimaging:bigguy|Big Guy]]. | + | {{ :confocal_pinhole.png?350|}} The Zeiss LSM 710 is a [[https://www.youtube.com/watch?v=rTkjQ6Q0p5I|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 cage incubator and a stage top incubator for temperature control and CO2 supply. The stage is motorized so imaging is possible in multiple stage positions or tiled acquisitions. Its scanning unit has a spectral recycling loop, two sensitive PMT detectors and a filter-free spectral separation module that can be continuously set over the entire wavelength range (i.e. you can specify which wavelength range you want to detect). It is equipped with lasers from violet to far red (405, 458, 488, 514, 561 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 and detection sensitivity are 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]], but image acquisition is slower. If you need to use a 594 nm laser or increased resolution and sensitivity, or are performing long time-lapse experiments in live samples (aqueous media) and need hardware focus control, check the [[zeiss_lsm_880|Zeiss LSM 880]] or [[zeiss_lsm_980|Zeiss LSM 980]] confocal point-scanning systems with Airyscan and Definite Focus. If your personal computer cannot handle all the data you collected, check out the [[colossus|Colossus]]. |
- | ===== Booking Rules ===== | + | {{:warning.png?25|}} Data files older than **3 months** will be automatically deleted on this system, please copy your data to the iMM server using the desktop link. |
- | The Zeiss LSM 710 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** | + | ===== Booking Rules ===== |
- | - 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) | + | - **Users** can book at maximum **8 hours per week** |
- | - The above rules do not apply for **weekends** and for work days **before 9:00** and **after 21:00** | + | - This usage restriction does not apply for **weekends** and for working 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 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 write their internal extension phone number in the booking Description field | ||
===== System components ===== | ===== System components ===== | ||
==== LASERs ==== | ==== LASERs ==== | ||
- | ^ Laser Unit ^ Wavelength ^ Maximum Power ^ Current Status ^ | + | ^ Laser Unit ^ Wavelength ^ Maximum Power ^ |
- | | Diode 405-30 | 405 nm | 30 mW | ok | | + | | Diode 405-30 | 405 nm | 30 mW | |
- | | Argon | 458, 488 and 514 nm | 25 mW | ok | | + | | Argon | 458, 488 and 514 nm | 25 mW | |
- | | DPSS 561-10 | 561 nm | 15 mW | ok | | + | | DPSS 561-10 | 561 nm | 15 mW | |
- | | HeNe633 | 633 nm | 5 mW | ok | | + | | HeNe633 | 633 nm | 5 mW | |
==== 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 | 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]] | |
- | | 20x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&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=440452-9903-000|EC Plan-Neofluar]] | Oil | 1.30 | 0.20 | | + | | 40x | EC Plan-Neofluar DIC | Oil | 1.30 | 0.20 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420462-9900-000/Objective-EC-Plan-Neofluar-40x-1.30-Oil-DIC-M27|420462-9900-000]] | |
- | | 63x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440762-9904-000|Plan-Apochromat]] | Oil | 1.40 | 0.19 | | + | | 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]] | |
**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 | | + | | 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 | | + | |
==== Filtersets (Ocular) ==== | ==== Filtersets (Ocular) ==== | ||
- | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Emission ^ | + | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ |
- | | 1 | Green | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=000000-1031-346&o=|FS38]] | 450-490 nm | 500-550 nm | | + | | 1 | Green | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=000000-1031-346&o=|FS38]] | 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 ===== | ||
* Turn on the ''MAIN SWITCH'' | * Turn on the ''MAIN SWITCH'' | ||
- | * Turn on the two switches: ''SYSTEMS/PC'' and ''COMPONENTS'' | + | * Turn on the ''SYSTEMS/PC'' switch |
- | {{:bioimaging:switches_710.jpg?100}} | + | {{switches_710.jpg?100}} |
- | * Turn the metal halide power source on (if needed) | + | * Turn on the metal halide fluorescent light source (if needed) |
- | {{:bioimaging:halide_710.jpg?80}} | + | {{halide_710.jpg?80}} |
- | * Turn the Ar laser on using the key (if needed) | + | * Turn on the Ar laser using the key (if needed) |
- | {{:bioimaging:laser_key_710.jpg?100}} | + | {{laser_key_710.jpg?100}} |
- | * Turn the computer on | + | * If you need CO2, open the CO2 valve |
- | * Login with your unit account | + | * Turn on the computer |
- | * Wait for network icon to change to ''connected'' state | + | * Login in to Windows with your Agendo credentials |
+ | * Turn on the ''COMPONENTS'' switch | ||
+ | {{switches_710.jpg?100}} | ||
+ | * Check that the computer is connected to an ''Unidentified network'' (click on the network icon in the taskbar) | ||
+ | {{::network_icon.png?200}} | ||
* Switch the Ar laser from ''idle'' ton ''run'' (don't change the ''current'') | * Switch the Ar laser from ''idle'' ton ''run'' (don't change the ''current'') | ||
- | {{:bioimaging:laser_run_710.jpg?100}} | + | {{laser_run_710.jpg?100}} |
- | * Start the ''ZEN 2009'' software | + | * Start the **ZEN Black** software |
===== Microscope Turn Off Procedure ===== | ===== Microscope Turn Off Procedure ===== | ||
- | If there's another user for this microscope in the next hour: | + | |
- | * Close the software, leave the lasers on an log off the computer. | + | If there is another user for this microscope in the __next hour__: |
- | * Clean up any objective were you used immersion oil. | + | * Close **ZEN**, leave the lasers on, 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 561 nm laser and incubator in the software (if used) | * Turn off the 561 nm laser and incubator in the software (if used) | ||
+ | * Close the LSM 710 CO2 valve (if used) | ||
* Switch the Ar laser from ''run'' to ''idle'' (if used) | * Switch the Ar laser from ''run'' to ''idle'' (if used) | ||
* Turn off the Ar laser using the key (if used) | * Turn off the Ar laser using the key (if used) | ||
- | * Clean up any objective were you used immersion oil. | + | * Clean up immersion objectives |
- | * Exit the software, shut down the computer. | + | * Close **ZEN**, shutdown the computer. |
- | * Turn off the metal halide lamp | + | * Turn off the metal halide fluorescence light source |
- | * Turn off the main switches ''SYSTEMS/PC'' and ''COMPONENTS'' | + | * Turn off the two switches: ''SYSTEMS/PC'' and ''COMPONENTS'' |
* Wait 5 min. for Ar laser cooldown | * Wait 5 min. for Ar laser cooldown | ||
- | * Turn off the ''MAIN SWITCH''. | + | * Turn off the ''MAIN SWITCH'' |
---- | ---- | ||
- | [[groups:bioimage:resources|Back to the Resources page]] | + | [[resources|Back to the Equipment page]] |