Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_510_meta_wiki.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lsm_510_meta_wiki.jpg" width=300></a>
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<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>Model</b>: LSM 510 META</a><br>
<b>Software</b>: LSM 510<br>
<b>Year</b>: 2006 <br>
<b>SN</b>: 2435000218 <br>
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&rarr; <a href="http://www.imm.fm.ul.pt/booking/old/month.php?user=ZEISSLSM5&rid=1"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss LSM 510 META Booking</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_510_meta_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 510 META Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_510_meta_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Zeiss LSM 510 META Usage Statistics</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_510_meta_repair"><img src="/facility/bioimaging/lib/exe/fetch.php?media=repair-icon.gif" width=16> Zeiss LSM 510 META Repair History</a>
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===== Microscope overview =====
The Zeiss LSM 510 META is a //confocal point-scanning microscope// able to generate high-resolution three-dimensional images of thick specimens. It is an inverted microscope suitable for live cell imaging and photobleaching experiments, equipped with a large size incubator for temperature control and a small stage incubator for CO2 supply. Its META detector can be used for spectral detection and subsequent linear unmixing of overlapping fluorescent stainings, however at a much lower sensitivity than its other two PMT detectors. A vast range of high power lasers are available from violet to far red (405, 458, 488, 514, 561, 594 and 633 nm excitation wavelengths) with matching extended filtersets. With this system you can perform optical sectioning of fluorescent samples which are too thick for a widefield system such as the [[zeiss_axiovert_200m|Zeiss Axiovert 200M]], at the cost of increasing photobleaching/photodamage and reducing sensitivity. Image resolution is higher than faster confocal systems such as a spinning disk or the [[zeiss_lsm_7_live|Zeiss LSM 7 Live]], but image acquisition is much slower. For higher sensitivity and reduced photobleaching/photodamage check the [[zeiss_lsm_710|Zeiss LSM 710]] confocal point-scanning system. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]].

Some examples...

{{liverbykirsten.gif?128x128}}{{drosophilabymarco.gif?128x128}}
  - Mouse liver infected with malaria parasite. XYZ 3D reconstruction, Specimen: Kirsten Hanson (2007), Instituto de Medicina Molecular, Lisboa, Portugal.
  - Drosophila hemocytes. XYt, 41 slices, Specimen: Marco Antunes (2007), Instituto de Medicina Molecular, Lisboa, Portugal.

===== Booking Rules =====
The Zeiss LSM 510 META 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**
  - 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)
  - The above rules do not apply for **weekends** and for work 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]]**.
  - 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 =====

==== LASERs ====
^  Laser Unit  ^  Wavelength  ^  Maximum Power  ^  Current Status  ^
|  Diode 405-30  |  405 nm  |  50 mW  |  ok  |
|  Argon/2  |  458, 488 and 514 nm  |  45 mW  |  ok  |
|  DPSS 561-10  |  561 nm  |  15 mW  |  **not working**  |
|  HeNe594  |  594 nm  |  2 mW  |  ok  |
|  HeNe633  |  633 nm  |  5 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=420340-9901-000|EC Plan-Neofluar]]  |  Dry  |  0.30  |  5.2  |
|  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  |
|  40x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=421767-9970-000|C-Apochromat]]  |  Water  |  1.20  |  0.28  |
|  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  |

**Upon request:**

^  Magnification   ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
|  5x  |  [[https://www.micro-shop.zeiss.com/?s=571974962233f6&l=en&p=uk&f=o&a=v&m=s&id=420330-9901-000|EC Plan-NeoFluar]]  |  Dry  |  0.16  |  18.5  |
|  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  |  [[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 (VIS) ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Emission  ^
|  1  |  none  |  none  |  none  |  none  |
|  2  |  Blue  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488001-9901-000&o=|FS01]]  |  359-371 nm  |  > 397 nm  |
|  3  |  Green  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488009-9901-000&o=|FS09]]  |  450-490 nm  |  > 515 nm  |
|  4  |  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  |
|  5  |  Green + Red  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488023-0000-000&o=|FS23]]  |  475-495 + 540-552 nm  |  515-530 + 580-630 nm  |

==== Filter and Dichroic sets ====
|  {{zeissmetaconfig.jpg?120}}  |  {{zeissmetachannel1.jpg?120}}  |  {{zeissmetachannel2.jpg?111}}  |  {{zeissmetachanneldic.jpg?120}}  |  {{zeissmetamaindic.jpg?100}}  |  {{zeissmetameta.jpg?160}}  |
|  Main configuration window  |  Emission filters for channel 1  |  Emission filters for channel 2  |  Channel separation dichroic  |  Laser separation dichroic  |  META channels  |

===== Microscope Turn On Procedure =====
  * Turn on the two main switches: ''SYSTEM/PC'' and ''COMPONENTS''.
{{switches.jpg?100|}}
  * Turn the metal halide power source on.
{{excite.jpg?100|}}
  * Turn the computer on
  * Login with your unit account
  * Wait for network icon to change to ''connected'' state
  * Start the ''LSM 510'' software
  * If you need temperature control, turn on the warm air heating...
{{heating.jpg?100|}}
  * ... and set the correct temperature on the control box
{{heating2.jpg?100|}}
  * If you need CO2, open the valve and turn on CO2 supply on the control box
{{co2.jpg?100|}}

===== 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. 
  * Clean up any objective were you used immersion oil.
  * **Make sure that there really is another user going to use the microscope.**
Else:
  * Clean up any objective were you used immersion oil.
  * Turn off the lasers in the software and shut down the computer.
  * Turn off the metal halide power source.
  * Wait 5 min. for laser cooldown.
  * Turn off the two main switches ''SYSTEM/PC'' and ''COMPONENTS''.

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