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zeiss_lsm_880 [2016/04/05 00:18]
bioimaging
zeiss_lsm_880 [2024/01/28 11:47] (current)
bioimaging [Microscope overview]
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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_2022.png"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_lsm_880_wiki_2022.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">​ZEISS Microscopy</​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>
 +&nbsp <br>
 +Data will be deleted after: <b>1 month</​b>​
 &nbsp <br> &nbsp <br>
 &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>​ 
- <​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>​
 </td> </td>
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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]]. +{{  :​confocal_pinhole.png?​340}} ​The Zeiss LSM 880 with Airyscan is a [[https://www.youtube.com/​watch?​v=rTkjQ6Q0p5I|point-scanning ​confocal ​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]] ​with 32 concentrically arranged GaAsP detection elements ​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 4x, 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 =====+
  
-==== LASERs ==== +  * [[https://​www.youtube.com/​watch?​v=sGTNBXq8omg|ZEISS Webinar: LSM 880 with Airyscan ​Revolutionize Your Confocal Imaging]] 
-^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^ +  ​* [[https://​www.youtube.com/​watch?​v=VuxgaKDa_DU|ZEISS Webinar: Fast Mode for ZEISS LSM 880 with Airyscan]]
- ​Sapphire 488-100  |  488 nm  |  100 mW  |  ok  | +
-|  DPSS 561-40 ​ |  561 nm  |  40 mW  |  ok  |+
  
-==== Objectives ==== +<​html>​ 
-^  Magnification ​ ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^ +<p align="​center"><​iframe width="​420"​ height="​236"​ src="https://​www.youtube.com/embed/​QjKrLTHGnc8"​ title="​YouTube video player"​ frameborder="​0"​ allow="​accelerometer;​ autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture"​ allowfullscreen><​/iframe> <iframe width="​420"​ height="​236"​ src="https://​www.youtube.com/embed/​ln6VUmLzLLc"​ title="​YouTube video player"​ frameborder="​0"​ allow="​accelerometer;​ autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture"​ allowfullscreen><​/iframe><​/p> 
-|  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  | +</html>
-|  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) ====+{{:​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.
  
-^  Position ​ ^  Filterset ​ ^  Reference ​ ^  Excitation ​ ^  Emission ​ ^ +===== Booking Rules ===== 
-|  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  | +  ​- ​**Users** can book at maximum **8 hours per week** 
-|  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  | +  ​- This usage restriction does not apply for **weekends** and for working days **before 9:00** and **after 21:00** 
- 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  | +  - Exceptions to these rules require **approval from [[joserino@medicina.ulisboa.pt|José Rino]]**.
-Excitation filter has been removed+
  
-==== Filter and Dichroic sets ==== +===== System components =====
-|  {{zeiss5livefilters0.jpg?​100}} ​ |  {{zeiss5livefilters1.jpg?​100}} ​ | +
-|  Main configuration ​ |  Emission filters ​ |+
  
-===== Microscope Turn On Procedures =====+==== LASERs ​==== 
 +^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^ 
 +|  Diode 405-30 ​ |  405 nm  |  30 mW  | 
 +|  Argon  |  458, 488 and 514 nm  |  25 mW  | 
 +|  DPSS 561-20 ​ |  561 nm  |  20 mW  | 
 +|  HeNe594 ​ |  594 nm  |  2 mW  | 
 +|  HeNe633 ​ |  633 nm  |  5 mW  |
  
-==== Normal Operation ​====+==== Objectives ​==== 
 +^  Magnification ​  ​^ ​ Model  ^  Immersion ​ ^  NA  ^  WD (mm)  ^  Reference ​ ^ 
 +|  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]] ​ | 
 +|  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  |  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]] ​ | 
 +|  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]] ​ |
  
-  * The system must be turned on **30 min** before usage. 
-    * 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 ====+**Upon request:**
  
-  ​* The system must be turned on **2 hours** before usage (in order to properly warm up scanning mirrors). +^  Magnification ​  ​^  Model  ^  Immersion ​ ^  NA  ^  WD (mm ​^ ​ Reference ​ ^ 
-    * To start it turn on the two main switches: **SYSTEM/PC** and **COMPONENTS** +|  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]] ​ | 
-  * Login to the computer, wait for the network icon to change to ''​connected''​ state + 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]] ​ | 
-  * Start the **ZEN 2009** software. +==== Filtersets (Ocular) ====
-**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 ​===== +^  Position ​ ^  Filterset ​ ^  Reference ​ ^  Excitation ​ ^  Dichroic ​ ^  Emission ​ ^ 
-If there'​s another user for this microscope in the next two hours: +|  2  |  Cyan  |  [[https://​www.chroma.com/​products/​sets/​49001-et-ecfp|49001]] ​ |  426-446 nm  |  455 nm  |  460-500 nm  | 
-  * Exit the **ZEN 2009** software, leave the lasers on and log off the computer.  +|  3  |  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  | 
-  * Clean up immersion objectives. + 4  |  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  | 
-  * **Make sure that there really is another user going to use the microscope.** + 5  |  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  |
-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: ​=====+===== 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 with your Agendo credentials 
 +  * Turn on the ''​COMPONENTS''​ switch 
 +  * Start the **ZEN Black** software
  
-  ​Please contact ​the [[about|Bioimaging Facility]] (Ext47316). The ablation system requires specific training even for users that are already trained ​on the LSM 7 Live itself.+===== 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 minfor Ar laser to cool down** 
 +  * Turn off the ''​MAIN SWITCH''​
  
 ---- ----
- +[[resources|Back to the Equipment ​page]]
-[[resources|Back to the Resources ​page]]+
zeiss_lsm_880.1459808308.txt.gz · Last modified: 2016/04/05 00:18 by bioimaging