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AdvancedImagingUIC

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Zeiss Stereo Lumar | Reservations

  • Stereo Lumar

  • The Zeiss' Stereo Lumar is a flurescence capable and fully automated stereoscope with an increased focusing range, which allows imaging of whole small animals (eg Zebrafish e c elegans), as well as cell cultures in petri dishes. Magnifications range is equivalent to ~1-12x, with variable zoom.


    Location    : Bartolomeu Dias Wing

    Microscope: Zeiss SteREO Lumar.V12

    Year Installed: 2011

    Optics: Fluorescence and off-axis illumination

    Cameras: Hamamatsu Orca-Flash (2.8MPx) and IDS uEye UI-3370CP (4.19MPx)

    Operating conditions: Room Temperature

    Options: None

    Keywords:

Suggestion for description in "Materials and Methods":

Images were acquired on Zeiss STereo LUMAR stereoscope, equiped with a Hamamatsu Orca-Flash 2.8 camera and GFP fluorescence filterset, controlled with the MicroManager v1.14 software.

Suggestion for "Acknowledgments":

We would like to thank the technical support of IGC's Advanced Imaging Facility (AIF-UIC), which is supported by the national Portuguese funding ref# PPBI-POCI-01-0145-FEDER-022122, co-financed by Lisboa Regional Operational Programme (Lisboa 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER) and Fundação para a Ciência e a Tecnologia (FCT; Portugal).

Installed Filtersets

Name
Excitation
Emission
CFP
436/20
480/40
YFP
500/20
535/30
GFP
470/40
525/50
TxRed
560/40
630/75

Spectra, lines and filters

Pixel size

Because this stereoscope has a continuous zoom it is important that you write down the magnification at which the images were acquired (as displayed on the microscope controler). TO calibrate the images in ImageJ or any other software you need to insert the correct pixel size by using the calculator below.

Hamamatsu Magnification: as displayed in controller LCD.

Calculated pixel size: 0 µmeter per 1 pixel

IDS Magnification: as displayed in controller LCD.

Calculated pixel size: 0 µmeter per 1 pixel

Setup

Turn on procedures

  1. Turn on the computer (if not on already) and log-in using your Agendo username and passowrd. Remember to book on the reservations system, even for very short 1 minute sessions.
  2. Turn on the mercury lamp power source.

    3. Fluorescent lamp house

  3. Turn the Transmitted light source if needed.

    4. Transmitted light source.

  4. Turn on the stereoscope control box.

    5. Stereoscope control box

  5. Turn on the camera power bar.
  6. Start the Micro-Manager software, available as a Desktop shortcut.

    7. Micro-Manager shortcut

  7. Set up your experiments on the stereoscope.
  8. Press "Live" button in MicroManager to open the fluorescence shutter.

Turn off procedures

  1. Check if there is anyone after you.

  2. Close all programs and Log-off from Windows.

    • If you skip this step, the next user will not be able to confirm his entry and he will be using the stereoscope with your name.
  3. Turn off the camera power bar.
  4. Turn off the stereoscope control box.
  5. If you used the mercury light source and there is no one using the stereoscope for the next 2 hours or you’re sure that the next user won’t use this light source, then turn it off.
  6. Clean the area.

Acquisition

  1. Make sure the stereoscope view is set to the CCD camera.

  2. On Micro-Manager press "Live" to get a live feed of your sample.

    live

  3. Readjust the focus on the stereoscope joystick until the image appears optimally sharp, and the exposure time on the software until you are satisfied with your image.
  4. Press "Snap" or "Snap to Album" to capture the image.
  5. On Micro-Manager save the image to your folder on D:\users.

Extra Info