Bioimaging Wiki

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<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>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>Software</b>: ZEN 2009 <br>
<b>Year</b>: 2007 <br>
<b>SN</b>: 2403000110 <br>
&nbsp <br>
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&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/new/12"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss LSM 7 Live Booking</a>
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&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>
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&rarr; <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>
&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>
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===== Microscope overview =====

{{  :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 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]].

{{info.gif?|}} Click on the image on the left to see the system beam path in higher detail

===== System components =====

==== LASERs ====
^  Laser Unit  ^  Wavelength  ^  Maximum Power  ^  Current Status  ^
|  Sapphire 488-100  |  488 nm  |  100 mW  |  ok  |
|  DPSS 561-40  |  561 nm  |  40 mW  |  ok  |

==== Objectives ====
^  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  |  3.1  |
|  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) ====

^  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  |
|  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 ====
|  {{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 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:
  * Turn off the lasers, wait 5 min for them to cool down.
  * 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: =====

  * 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.

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