Bioimaging Wiki

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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lightsheet_z1_map.jpg">Room P3-A-3</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/imaging-systems/lightsheet-z-1.html" target="_blank">Lightsheet Z.1</a><br>
<b>Software</b>: ZEN 2014 SP1 <br>
<b>Year</b>: 2016 <br>
<b>SN</b>: 2583000193 <br>
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&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/month.php?user=ZEISSLIGH&rid=129"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss Lightsheet Z1 Booking</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lightsheet_z1_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Zeiss Lightsheet Z1 Usage Statistics</a>
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===== Microscope overview =====

The Zeiss Lightsheet Z1 is a //light sheet fluorescence microscope// able to image optical sections of large samples at subcellular resolution and very fast rates, with almost no phototoxicity or bleaching. It splits fluorescence excitation and detection into two separate light paths, [[http://www.kcci.virginia.edu/sites/kcci.virginia.edu/files/lightshield.jpg|with the axis of illumination being perpendicular to the detection axis]]. 

{{:lightsheet.jpg?0x300|}}{{:lightsheet_z.jpg?0x300|}}

Since only a single thin section of the sample is illuminated by the light sheet, optical sectioning is achieved without a pinhole or image processing deconvolution. Light from the in-focus plane is collected by a sCMOS camera rather than pixel by pixel as in a point scanning laser confocal. This allows you to collect images faster and with less excitation light than you would with many other optical-sectioning microscopy techniques. You can image living samples in water or fixed samples where [[http://blogs.zeiss.com/microscopy/news/en/references-for-clearing-protocols/|tissue clearing]] has been performed. Samples are mounted not on coverslips but inside special chambers that provide heating and cooling. The system is equipped with lasers from green to far red (488, 561 and 638 nm excitation wavelenghts) and appropriate fluorescence emission filters. With this system you can image live drosophila and zebrafish samples or use tissue clearing to perform optical sectioning deep into large fluorescent samples such as tissue sections, brains, embryos, organs, spheroids or biopsies. If you need higher resolution and have a smaller and thinner sample, check instead a confocal microscope such as the spinning disk [[3i_marianas_sdc|3i Marianas SDC]], the //line scanning// [[zeiss_lsm_7_live|Zeiss LSM 7 Live]] or the point scanners [[zeiss_lsm_880|Zeiss LSM 880]] and [[zeiss_lsm_710|Zeiss LSM 710]]. A dedicated computer running [[https://www.arivis.com/en/imaging-science/arivis-vision4d|arivis Vision4D]] is available for working with multi-channel 2D, 3D and 4D images you generate in the system. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]].

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

==== LASERs ====
^  Laser Unit  ^  Wavelength  ^  Maximum Power  ^  Current Status  ^
|  Solid State 488  |  488 nm  |  30 mW  |  ok  |
|  Solid State 561 |  561 nm  |  20 mW  |  ok  |
|  Solid State 638  |  638 nm  |  75 mW  |  ok  |

==== Objectives (Illumination) ====

^  Magnification  ^  Model  ^  NA  ^
|  10x  |  Lightsheet Z.1 10x  |  0.20  |

==== Objectives (Detection) ====

^  Magnification   ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
|  20x  |  [[https://www.micro-shop.zeiss.com/?s=10396935ab3601&l=en&p=uk&f=o&a=v&m=s&id=421452-9700-000|W Plan-Apochromat]]  |  Water  |  1.00  |  2.4  |
|  20x  |  Clr Plan-Neofluar Corr nd=1.45  |  Clearing  |  1.00  |  5.6  |

==== Filtersets (Water) ====

^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  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  |  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  |  570-640 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  |  420-470 nm  |

===== Microscope Turn On Procedure =====
  * Turn on the ''MAIN SWITCH''
  * Turn on the two switches: ''SYSTEMS/PC'' and ''COMPONENTS''
{{zeiss_lsm_880_1.jpg?100}}
  * Turn the computer on
  * Login with your unit account
  * Wait for network icon to change to ''connected'' state
  * Start the **ZEN** software

===== Microscope Turn Off Procedure =====
If there's another user for this microscope in the next hour:
  * Close the software, leave the lasers on and log off the computer. 
  * Clean up any objective where you used immersion oil.
  * **Make sure that there really is another user going to use the microscope.**
Else:
  * Turn off the lasers on **ZEN**
  * Clean up any objective were you used immersion oil.
  * Exit the software, shut down the computer.
  * Turn off the main switches ''SYSTEMS/PC'' and ''COMPONENTS''
  * Wait 5 min. for Ar laser cooldown
  * Turn off the ''MAIN SWITCH''.

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