User Tools

Site Tools


zeiss_lsm_7_live

Differences

This shows you the differences between two versions of the page.

Link to this comparison view

Next revision
Previous revision
zeiss_lsm_7_live [2016/03/10 01:26]
bioimaging created
zeiss_lsm_7_live [2021/01/17 04:48]
bioimaging [If you want to use the UV laser ablation system:]
Line 6: Line 6:
 <a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_lsm_7_live_wiki.jpg"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_lsm_7_live_wiki.jpg"​ width=300></​a>​ <a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_lsm_7_live_wiki.jpg"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=zeiss_lsm_7_live_wiki.jpg"​ 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_7_live_map.jpg">​Room P2-B-42</​a>​ (<img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=telephone_icon.gif"​ width=15>​ 47246) ​<​br> ​+<​b>​Location</​b>: ​no longer available ​<​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.ticgroup.com.tw/​menu/​products/​sci/​lsms/​slsm/​lsm_7_live/​45-0067_elive.pdf"​ target="​_blank">​LSM 5 LIVE</​a><​br>​ <​b>​Model</​b>:​ <a href="​http://​www.ticgroup.com.tw/​menu/​products/​sci/​lsms/​slsm/​lsm_7_live/​45-0067_elive.pdf"​ target="​_blank">​LSM 5 LIVE</​a><​br>​
Line 14: Line 14:
 &nbsp <br> &nbsp <br>
 &nbsp <br> &nbsp <br>
-&rarr; <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 7 Live Booking</​a>​+&nbsp <​br>​ 
 +&rarr; <a href="​https://​imm.medicina.ulisboa.pt/​intranet/​booking/​12"><​img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=date.png"> ​ Zeiss LSM 7 Live Booking</​a>​
  <​br> ​  <​br> ​
 &rarr; <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>​ &rarr; <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>​
Line 21: Line 22:
 &nbsp </br> &nbsp </br>
 &rarr; <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>​ &rarr; <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>​ + </​p>​
-&rarr; <a href="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=bioimaging:​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>​ +
-&​nbsp ​</p>+
 </td> </td>
 </tr> </tr>
Line 29: Line 28:
 </​html>​ </​html>​
  
-===== System ​overview ===== +===== Microscope ​overview =====
-The 3i Marianas SDC //spinning disk confocal microscope//​ is a fast imaging system which provides a trade-off between confocality,​ resolution and speed. It is an inverted microscope ideal for live cell applications which require fast acquisition speeds rather than high resolution images. The scanning unit achieves confocality by directing light through a [[http://​zeiss-campus.magnet.fsu.edu/​tutorials/​spinningdisk/​yokogawa/​yokogawatutorialfigure1.jpg|spinning disk with many small pinholes]]. Images are then acquired with a sensitive EMCCD which allows for very small exposure times but is limited in resolution to 512x512 pixels. The stage is motorized and furthermore equipped with a piezo for Z displacement so fast 4D imaging is possible in multiple stage positions. The system is also equipped with a large size incubator for temperature control and a small stage incubator for CO2 supply. 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_wf|Zeiss Cell Observer WF]]. Even though its resolution 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]], it is much faster than these two systems. If you need to use an upright microscope, check the [[zeiss_lsm_7_live|Zeiss LSM 7 Live]] fast confocal system instead. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. ​+
  
-  * **Microscope**[[http://​www.zeiss.com/microscopy/en_de/products/light-microscopes/​axio-observer-for-biology.html|Zeiss ​Axio Observer]] +{{  ​:lsm7live_lighpath.jpg?​0x320|}} 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 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 interestThe 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 linesIf your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. 
-  * **Confocal scanner**: ​[[http://​www.andor.com/​microscopy_systems/​peripherals/​confocal_scanners/​|Yokogawa CSU-x1]] + 
-  * **Camera**: ​[[http://www.photometrics.com/​products/​emccdcams/​evolve/​512.php|Evolve 512 EMCCD]]+Click on the image on the left to see the system beam path in higher detail
  
 ===== System components ===== ===== System components =====
Line 40: Line 38:
 ==== LASERs ==== ==== LASERs ====
 ^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^ ^  Laser Unit  ^  Wavelength ​ ^  Maximum Power  ^  Current Status ​ ^
-|  ​Laserstack ​488  |  488 nm  |  100 mW  |  ok  | +|  ​Sapphire ​488-100  ​| ​ 488 nm  |  100 mW  |  ok  | 
-|  ​Laserstack ​561  |  561 nm  |  ​100 mW  |  ok  | +|  ​DPSS 561-40  ​| ​ 561 nm  |  ​40 mW  |  ok  |
-|  Laserstack 640  |  640 nm  |  100 mW  |  ok  |+
  
 ==== Objectives ==== ==== Objectives ====
-^  Magnification ​  ​^ ​ Model  ^  Type  ^  NA  ^  WD (mm)  ^ 
-|  20x  |  [[https://​www.micro-shop.zeiss.com/?​s=129353259d52f12&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420650-9901-000|Plan-Apochromat]] ​ |  Dry  |  0.8  |  0.55  | 
-|  63x  |  [[https://​www.micro-shop.zeiss.com/?​s=3407786617d462&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420782-9900-000|Plan-Apochromat]] ​ |  Oil  |  1.40  |  0.19  | 
-|  100x  |  [[https://​www.micro-shop.zeiss.com/?​s=129353259d52f12&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420790-9901-000|Plan-Apochromat]] ​ |  Oil  |  1.40  |  0.17  | 
- 
-**Upon request:** 
 ^  Magnification ​ ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^ ^  Magnification ​ ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
-|  ​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/ ​| ​ 0. ​|  ​0.21  | +|  ​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  | 
-|  ​40x  ​| ​ [[https://​www.micro-shop.zeiss.com/?​s=129353259d52f12&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=420660-9970-000|Plan-Apochromat]] ​ |  ​Dry  ​|  ​0.95  ​| ​ 0.25  | +|  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. | 
-==== Emission ​Filtersets ==== +|  ​20x  ​| ​ [[https://​www.micro-shop.zeiss.com/?​s=3407786617d462&​l=en&​p=uk&​f=o&​a=v&​m=s&​id=421452-9900-000|Plan-Apochromat]] ​ |  ​Water dipping ​ ​|  ​1.00  |  1.8  | 
-^  ​Setting ​ ​^  ​Transmission ​ ^ +|  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  | 
-|  ​1: 445  ​|  ​422-477 nm  + 
-|  ​2525  ​|  ​510-540 nm  | + 
-|  3: 617  ​|  ​580-653 nm  +==== Filtersets ​(Ocular) ​==== 
-|  ​4692  ​|  ​672-712 nm  | + 
-|  ​5: QUAD  ​|  ​440+521+607+700 nm  +^  ​Position ​ ​^  ​Filterset ​ ^  Reference ​ ^  Excitation ​ ^  Emission ​ ^ 
-|  ​6Block  ​|  ​No Transmission ​ | +|  ​ ​|  ​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  | 
-|  ​7: Block  ​|  ​No Transmission ​ | +|  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  | 
-|  ​8: Block  ​|  ​No Transmission ​ | +|  ​ ​|  ​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  | 
- 9Empty  ​|  ​Empty  | +* Excitation filter has been removed 
-|  ​10445   422-477 nm  |+ 
 +==== Filter and Dichroic sets ==== 
 +|  ​{{zeiss5livefilters0.jpg?​100}} ​ ​|  ​{{zeiss5livefilters1.jpg?​100}} ​ | 
 +|  ​Main configuration ​ ​|  ​Emission filters ​ | 
 + 
 +===== Microscope Turn On Procedures ===== 
 + 
 +==== Normal Operation ==== 
 + 
 +  * 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 ==== 
 + 
 +  * 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 On Procedure ​===== +===== Microscope Turn Off procedures ​===== 
-  * Check that the main power supply switch is on (should be on by default) +If there'​s another user for this microscope in the next two hours: 
-{{marianas_power1.jpg?|}} +  * Exit the **ZEN 2009** software, leave the lasers ​on and log off the computer. ​ 
-  * Turn on the fluorescence lamp (if needed) +  * Clean up immersion objectives
-{{marianas_power5.jpg?|}} +  * **Make sure that there really is another user going to use the microscope.** 
-  * Turn on the secondary power supply switch +Else: 
-{{marianas_power2.jpg?|}} +  * Turn off the lasers, wait 5 min for them to cool down
-  * Turn on the power switch distributor (green) and the component power switches (red) **one at a time** +  * Exit the **ZEN 2009** software and shut down the computer.  
-{{marianas_power3.jpg?|}} +  * Clean up immersion objectives
-  * Turn on the Yokogawa spinning disk scanner (using the key) +  * Turn off the metal halide lamp
-{{marianas_power4.jpg?|}} +  * Turn off the microscope
-  * Turn on incubation and CO2 on the touchscreen (if needed) +  * Turn off the two main switches: ''​SYSTEM/​PC''​ and ''​COMPONENTS''​
-{{marianas_power6.jpg?|}} +
-  * Turn on the lasers you need to use (using the key and the power switches) +
-{{marianas_power7.jpg?|}} +
-  * Turn on the computer +
-  * Wait 2 min to allow for system components detection +
-  * Start the SlideBook software+
  
-**Warning**: ​If you need to change ​the stage adaptor, please contact the <​html><​a href="/​facility/​bioimaging/​doku.php?​id=about">​Bioimaging Facility</​a>​ (<​b>​imm-bioimaging@medicina.ulisboa.pt</​b>​ | <img src="/​facility/​bioimaging/​lib/​exe/​fetch.php?​media=telephone_icon.gif"​ width=15> 47316</​html>​)+===== If you want to use the UV laser ablation system: ​=====
  
-===== Microscope Turn Off Procedure ===== +  ​Please contact ​the [[about|Bioimaging Unit]] (Ext: 47316). The ablation ​system ​requires specific training even for users that are already trained on the LSM 7 Live itself.
-  ​Remove your sample from the stage/piezo holder +
-  * Exit the SlideBook software +
-  * Switch off the computer +
-  * Turn off all system ​components (follow ​the Turn On Procedure in reverse)+
  
 ---- ----
  
-[[resources|Back to the Resources ​page]]+[[resources|Back to the Equipment ​page]]
zeiss_lsm_7_live.txt · Last modified: 2021/01/17 04:48 by bioimaging