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zeiss_lsm_880 [2016/04/05 00:18]
bioimaging
zeiss_lsm_880 [2022/01/20 13:34]
bioimaging [Microscope Turn On Procedure]
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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">
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-<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>​
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-<​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>:​ <a href="https://www.youtube.com/watch?v=sGTNBXq8omg">​LSM 880 with Airyscan</​a> ​(click for Zeiss webinar) <​br>​ 
-<​b>​Software</​b>:​ ZEN 2.(black) <br>+<​b>​Nickname</​b>:​ "​880"​<​br>​ 
 +<​b>​Software</​b>:​ ZEN 2.3 SP1 (black ​edition) <br>
 <​b>​Year</​b>:​ 2016 <br> <​b>​Year</​b>:​ 2016 <br>
 <​b>​SN</​b>:​ 2811000267 <br> <​b>​SN</​b>:​ 2811000267 <br>
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 &nbsp <br> &nbsp <br>
 &nbsp <br> &nbsp <br>
-&rarr; <a href="​https://​imm.medicina.ulisboa.pt/intranet/booking/​month.php?user=ZEISSLSM8&​rid=128"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=date.png"> ​ Zeiss LSM 880 Booking</​a>​+&rarr; <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>​
  <​br> ​  <​br> ​
 &rarr; <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>​ &rarr; <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>​  <​br>​
 &rarr; <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>​ &rarr; <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>​
-&nbsp </br> 
-&rarr; <a href="/​facility/​bioimaging/​doku.php?​id=zeiss_lsm_880_repair"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=repair-icon.gif"​ width=16>​ Zeiss LSM 880 Repair History</​a>​ 
  </​p>​  </​p>​
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 ===== Microscope overview ===== ===== Microscope overview =====
  
-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 ​specially ​suitable for live cell imaging and photobleaching experiments,​ equipped with a large size incubator for temperature control and CO2 supply. 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 ​for single photon counting) as well as a GaAsP detector. ​ 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]]. +{{  :​airyscan_beampath.png?​400}} ​The Zeiss LSM 880 with Airyscan is a [[https://imm.medicina.ulisboa.pt/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=confocal.png|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 cage incubator and a stage top 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). ​ 
-===== System components =====+{{::​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]].
  
-==== LASERs ==== +{{:​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
-^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^ +
-|  Sapphire 488-100 ​ |  488 nm  |  100 mW  |  ok  | +
-|  DPSS 561-40 ​ |  561 nm  |  40 mW  |  ok  ​|+
  
-==== Objectives ==== +{{:warning.png?25|}} Data files older than **month** will be automatically deleted on this system, please copy your data to the iMM server using the desktop link.
-^  Magnification ​ ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^ +
-|  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. | +
-|  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  |  [[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  |  [[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  | +
-|  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  |+
  
-==== Filtersets (Ocular) ​====+===== Additional information =====
  
-^  Position ​ ^  Filterset ​ ^  Reference ​ ^  Excitation ​ ^  Emission ​ ^ +  * [[https://​www.youtube.com/watch?v=sGTNBXq8omg|ZEISS WebinarLSM 880 with Airyscan ​Revolutionize Your Confocal Imaging (video)]] 
-|  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  | +  * [[https://​www.youtube.com/watch?v=VuxgaKDa_DU|ZEISS Webinar: Fast Mode for ZEISS LSM 880 with Airyscan (video)]]
-|  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  | +
- 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  | +
-* Excitation filter has been removed+
  
-==== Filter and Dichroic sets ==== +===== Booking Rules ===== 
- {{zeiss5livefilters0.jpg?​100}} ​  {{zeiss5livefilters1.jpg?​100}} ​ | +  ​- **Users** can book at maximum **8 hours per week** 
-|  Main configuration ​ |  Emission filters  ​|+  - This usage restriction does not apply for **weekends** and for working days **before 9:00** and **after 21:00** 
 +  - Exceptions to these rules require **approval from [[joserino@medicina.ulisboa.pt|José Rino]]**.
  
-===== Microscope Turn On Procedures ​=====+===== System components ​=====
  
-==== Normal Operation ​====+==== LASERs ​==== 
 +^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^ 
 +|  Diode 405-30 ​ |  405 nm  |  30 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  |
  
-  ​* The system must be turned on **30 min** before usage. +==== Objectives ==== 
-    * To start it turn on the two main switches''​SYSTEM/PC''​ and ''​COMPONENTS''​ +^  Magnification ​  ​^  Model  ^  Immersion ​ ^  NA  ^  WD (mm)  ^  Reference ​ ^ 
-{{7live_switches.jpg?100|}} +|  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]] ​ | 
-  * Login to the computer, wait for the network icon to change to ''​connected''​ state +|  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]] ​ | 
-  ​* Start the **ZEN 2009** software. + 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]] ​ | 
-**After the 30 min:** +|  40x  |  LD C-Apochromat Corr  |  Water  |  1.10  |  0.62  |  [[https://​www.micro-shop.zeiss.com/​en/​de/​shop/​objectives/​421867-9970-000/​Objective-LD-C-Apochromat-40x-1.1-W-Corr-M27|421867-9970-000]]  ​
-  ​* Turn on the metal halide lamp (white box to the left of the microscope)+ 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]] ​ |
-{{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). +**Upon request:**
-    * 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 ===== + Magnification ​  ​^ ​ Model  ^  Immersion ​ ^  NA  ^  WD (mm)  ^  Reference ​ ^ 
-If there'​s another user for this microscope in the next two hours: + 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]] ​ | 
-  * Exit the **ZEN 2009** software, leave the lasers on and log off the computer.  + 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]] ​ | 
-  * Clean up immersion objectives. +==== Filtersets (Ocular) ====
-  * **Make sure that there really is another user going to use the microscope.** +
-Else: +
-  ​* Turn off the lasers, wait 5 min for them to cooldown. +
-  * Exit the **ZEN 2009** software and shut down the computer +
-  * Clean up immersion ​objectives. +
-  ​* Turn off the metal halide lamp. +
-  * Turn off the microscope. +
-  * Turn off the two main switches**SYSTEM/PC** and **COMPONENTS**+
  
-===== If you want to use the UV laser ablation system: =====+^  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  |  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 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  |  395 nm  |  420-470 nm  |
  
-  ​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.+===== Microscope Turn On Procedure ===== 
 +  ​Turn on the ''​MAIN SWITCH''​ 
 +{{zeiss_lsm_880_1.jpg?​100}} 
 +  * Turn on the ''​SYSTEMS/​PC''​ switch 
 +  * Turn on the computer 
 +  * Login in to Windows ​(Bioimaging User) 
 +  * Turn on the ''​COMPONENTS''​ switch 
 +  * Start the **ZEN Black** software
  
-----+===== Microscope Turn Off Procedure ===== 
 +If there is another user for this microscope in the __next hour__: 
 +  * Exit the ZEN software, leave the lasers on 
 +  * Log off the computer 
 +  * Clean up immersion objectives 
 +Else: 
 +  * Turn off the lasers on **ZEN** 
 +  * Clean up immersion objectives 
 +  * Exit the ZEN software 
 +  * Shutdown the computer 
 +  * Turn off the two switches ''​SYSTEMS/​PC''​ and ''​COMPONENTS''​ 
 +  * **Wait 5 min. for Ar laser to cool down** 
 +  * Turn off the ''​MAIN SWITCH''​
  
-[[resources|Back to the Resources ​page]]+---- 
 +[[resources|Back to the Equipment ​page]]
zeiss_lsm_880.txt · Last modified: 2024/01/28 11:47 by bioimaging