Mizar Tilt User Manual
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Mizar Tilt User Manual

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Tilt User's Guide
Tilt
User's Guide
v1.0
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www.mizarimaging.com

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Summary of Contents for Mizar Tilt

  • Page 1 Tilt User’s Guide Tilt User’s Guide v1.0 www.mizarimaging.com...

  • Page 2: Table Of Contents

    Advanced Alignment and Installation Installation Aligning Tilt Optical Components Tips for Mounting Samples Recommended Specimen Chambers Assembling Mizar Cube Chambers Troubleshooting 7 MBL Street Lillie 220 Woods Hole, MA 02543 +1.508.939.7500 info@mizarimaging.com www.mizarimaging.com www.mizarimaging.com © 2019 Mizar Imaging, LLC. All Rights Reserved.

  • Page 3: Warnings And Safety

    This product is designed to be completely safe when used as intended. Any other use of the Tilt not outlined in this manual — including modifications and/or disabling safety fea- tures — may result in serious injury or property damage. In order to ensure safe operation, take care to review the following safety information before using the Tilt.

  • Page 4: Warnings

    Before operating the Tilt or turning on the laser source, ensure that the optical fiber carry- ing the laser light is securely fixed to both the Tilt and the laser launch. Check that all safety features included with the Tilt are in place.

  • Page 5: Safety Features

    Tilt User’s Guide Safety Features In order to reduce the risk of injury due to laser exposure, the Tilt is equipped several safety features. These include, a laser shield (I), a light trap (II), locking screws (III), and a fiber adapter cover (IV).

  • Page 6: How The Tilt Works

    The design of most LSFM/SPIM microscopes involve the use of two objectives, the orientation of which limits the numerical aperture (NA) of the detection objective. The Tilt from Mizar Imaging gives researchers the freedom to use LSFM/SPIM with high resolution, high-NA objectives.

  • Page 7: Getting To Know Your Tilt

    Tilt (Fig A). 1 – FRONT HEIGHT ADJUSTMENT (GREEN) The Green knob adjusts the height of the front side of the Tilt (Fig B), effectively sweeping the sheet up and down above the objective. Adjust the Green knob to position the light sheet vertically at the focal point of your objective.

  • Page 8: Getting To Know Your Tilt

    Tilt User’s Guide Getting to Know Your Tilt www.mizarimaging.com...

  • Page 9: Adjustment Tips

    1-4 mm. Zero the included inclinometer to the stage and then place it on one of the Tilt sled arms. Turn the (Pink) Rear Height Adjustment knob (Fig A-3) until the inclinometer reads the desired Tilt angle (~3.3-3.4°).

  • Page 10: How To Image

    • With the laser on low power (~2 mW) take a piece of lens paper (or a lint-free wipe) and hold it over the front lens cover where the four light sheets emerge from the Tilt optic assembly.
  • Page 11 • Use the (Green) knob to set the lightsheet to the centermost “dim” spot. • Turn the (Blue) knob to move the Tilt assembly either forward or back — in the direction that makes the image on screen brighter.

  • Page 12: Advanced Alignment And Installation

    Tilt. 1.3 Affix the Tilt stage Place the Tilt stage atop your stand and affix it to the stage adapters or through the spacers using the screws from your existing microscope stage. 1.4 Install the Tilt optical assembly Place the Tilt optical assembly on top of the three adjustable posts (above the Green and Pink knobs) on your Tilt stage.

  • Page 13: Aligning Tilt Optical Components

    WARNING: Aligning optical components should only be carried out by installer or certi- fied engineer. Your Tilt will be properly aligned upon its initial installation, and only on rare instances will it become necessary to perform a realignment. If you feel your Tilt optical components require realigning call our trained scientists at +1.508.939.7500.
  • Page 14 To collimate the beam: Couple a low powered alignment laser (“Visual Fault Locator” or similar) to the back of the Tilt. Make sure the fiber and fiber coupler are matched (angled or straight). Alterna- tively, use a laser launch set to <2mW.
  • Page 15 Installation To align the mirrors: Open the top lid of the Tilt and hang the alignment target over the front edge, blocking the path to the lens. Make sure the bottom of the target is flush with the bottom surface of the Tilt enclosure and center the target in the window.  ...
  • Page 16 Tilt User’s Guide Advanced Alignment and Installation Mirror Alignment Tips: • The best way to get a sense of how the alignment works is to adjust the knobs and monitor how the adjustment affects the position and shape of your beam - both at the target before the lens and the sheets after the lens.

  • Page 17: Tips For Mounting Samples

    For instance, if you immobilize your sample in 0.5% agarose, then do that as well for imaging with the Tilt; if you do some sort of trick to bring your sample as close to the cover slip as possible, such as glue, then use that as well with the Tilt; etc.

  • Page 18: Recommended Specimen Chambers

    Tilt User’s Guide Recommended Specimen Chambers The key to successful imaging with the Tilt is an imaging chamber with flat, optically clear sides to allow an unobstructed path for the lightsheet. Currently there are two commercial options we routinely use.

  • Page 19: Assembling Mizar Cube Chambers

    Spread a thin layer of vacuum grease on the inside of a petri dish, or similar flat surface. Using a gloved hand to avoid fingerprints, firmly press a Mizar Cube into the thin layer of grease; inspect the bottom of the Mizar Cube for complete coverage with the grease.
  • Page 20 Mizar Cube. The assembled Mizar Cube chamber is now ready to loaded with your sample. Alterna- tively, a sample can be immobilized on a 22x40mm coverslip prior to sealing the Mizar Cube chamber around it and filling with media. www.mizarimaging.com...

  • Page 21: Troubleshooting

    Centering the lightsheet in your field of view is more critical at low magnifi- cation, as the entire 300μm “waist” will be in use. Make sure the Tilt angle is optimal at 3.3° and use the Blue knob to better center the lightsheet to your image.
  • Page 22 Tilt User’s Guide 7 MBL Street Lillie 220 Woods Hole, MA 02543 +1.508.939.7500 info@mizarimaging.com www.mizarimaging.com www.mizarimaging.com...

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