GIMM Bioimaging Wiki

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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_lightsheet_z1_map_new_p2a24.png">[Lisbon] Room P2-A-24</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47219) <br> 
<b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">ZEISS Microscopy</a><br>
<b>Model</b>: <a href="https://www.youtube.com/watch?v=vKHQudCKVGc" target="_blank">Lightsheet Z.1</a> (click for Zeiss webinar)<br>
<b>Nickname</b>: "Lightsheet"<br>
<b>Software</b>: ZEN 2014 SP1 <br>
<b>Year</b>: 2016 <br>
<b>SN</b>: 2583000193 <br>
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Data will be deleted after: <b>3 months</b>
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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 Z.1 Usage Statistics</a>
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===== Microscope overview =====

{{  :lightsheet_z.jpg?0x380|}} The Zeiss Lightsheet Z.1 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. [[https://www.zeiss.com/content/dam/Microscopy/Products/imaging-systems/lattice-lightsheet/lattice-lightsheet-7/zeiss-lattice-lightsheet-7-beam-splitting.svg|It splits fluorescence excitation and detection into two separate light paths]], with the axis of illumination being perpendicular to the detection axis. 
[[https://imm.medicina.ulisboa.pt/facility/bioimaging/lib/exe/fetch.php?media=lightsheet_illumination.jpg|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 [[https://imm.medicina.ulisboa.pt/facility/bioimaging/lib/exe/fetch.php?media=en_poster_overview-clearing-methods_rel-2.0.pdf|tissue clearing]] has been performed. Samples are mounted not on coverslips but inside [[https://imm.medicina.ulisboa.pt/facility/bioimaging/lib/exe/fetch.php?media=lightsheet_chambers.jpg|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. A dedicated computer running [[https://www.arivis.com/|arivis]] is available for working with multi-channel 2D, 3D and 4D images you generate in the system. 

{{:info.png?25|}} Click on the image on the right to see the system beam path in higher detail

{{:warning.png?25|}} Data files older than **3 months** will be automatically deleted on this system, please copy your data to the GIMM server using the desktop link.

===== Additional information for sample preparation =====

  * **Sample Size**: up to 1 x 1 x 2 cm
  * **Penetration depth**: up to 5.6 mm

    * [[https://imm.medicina.ulisboa.pt/facility/bioimaging/lib/exe/fetch.php?media=en_poster_overview-clearing-methods_rel-2.0.pdf|Clearing Methods in Microscopy (poster)]] 
    * [[https://hcbi.fas.harvard.edu/files/lightsheetz1_sample-preparation_zeiss.pdf| Sample Preparation for Light Sheet Microscopy (white paper)]]
    * [[https://asset-downloads.zeiss.com/catalogs/download/mic/b91bc689-66ab-4d85-b3f6-c31ebea22ba7/EN_wp_Lightsheet-Z.1_Expanding-Clearing-Solutions.pdf| Improved Imaging of Cleared Samples (application note)]]
  * [[https://www.youtube.com/watch?v=Ki1wtoXIkis|Sample Preparation for Whole Organisms, Plants, and 3D Cell & Tissue Cultures (video)]]
  * [[https://www.youtube.com/watch?v=Ql7T7LrvI_I|ZEISS Microscopy How-to: Mount cleared samples in Lightsheet Z.1 (video)]]
  * [[https://www.youtube.com/watch?v=vKHQudCKVGc|ZEISS Webinar: In Vivo and Cleared Tissue Imaging with Light Sheet Fluorescence Microscopy (video)]]

===== Tutorials for Image Analysis with Arivis =====
  * [[https://www.arivis.com/lunchtime-academy|Lunchtime Academy – Getting started with arivis Vision4D]]

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

==== LASERs ====

^  Laser Unit  ^  Wavelength  ^  Maximum Power  ^
|  Blue  |  <fc #00f7ff>488</fc>  |  30 mW  |
|  Yellow-Green  |   <fc #c6ff00>561 nm</fc>  |  20 mW  |
|  Red  |  <fc #ff4200>638 nm</fc>  |  75 mW  |

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

^  Magnification  ^  Model  ^  NA  ^
|  <fc #fef042>10x</fc>  |  Lightsheet Z.1 10x  |  0.20  |

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

^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  <fc #0aa05c>20x</fc>  |  W Plan-Apochromat DIC 75mm  |  Water  |  1.00  |  2.4  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/421452-9700-000/Objective-W-Plan-Apochromat-20x-1.0-DIC-M27-75mm#|421452-9700-000]]  |
|  <fc #0aa05c>20x</fc>  |  Clr Plan-Neofluar Corr nd=1.45 85mm  |  Clearing  |  1.00  |  5.6  |  [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/421459-9970-000/Objective-Clr-Plan-Neofluar-20x-1.0-Corr-nd-1.45-M32-85mm|421459-9970-000]]  |

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

^  Filterset  ^  Emission  ^
|  GFP  |  505-545 nm  |
|  Cy3  |  575-615 nm  |
|  DRAQ5  |  LP 660 nm  |

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