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v2.0

by z mbstaff

Introduction

This guide will walk you through the startup process of the Nikon spinning disk confocal microscope, located at Irchel Campus, Room 42 F 66.

Please find more detailed information about this system in here.

  1. Start the  computer below the table. Start the microscope controller  below the optical table Start the spinning disk unit
    • Start the computer below the table.

    • Start the microscope controller below the optical table

    • Start the spinning disk unit

    • Start the both cameras

    • It is very important that the camera will be switched on after the computer is booted. Otherwise you risk the blue death of the computer!

  2. Follow this guide if you need to turn on environmental control for live cell imaging.
    • Follow this guide if you need to turn on environmental control for live cell imaging.

  3. Localize the water dispenser Check if there is enough water in the bottle. If not, find a bottle on the wall and fill the water dispenser.
    • Localize the water dispenser

    • Check if there is enough water in the bottle.

    • If not, find a bottle on the wall and fill the water dispenser.

    • If you see that there is almost no water left in the bottle, please let the ZMB staff know!

  4. Pres Ctrl + Alt + Del and log in with your CORE credentials
    • Pres Ctrl + Alt + Del and log in with your CORE credentials

  5. Star the NIS-Elements SW Star the software either in single or dual camera mode.
    • Star the NIS-Elements SW

    • Star the software either in single or dual camera mode.

  6. Open the black environmental control box to access the stage. Turn on the light in the box, if needed. Make sure that the objective is in the lowest position. If you are not sure, engage the escape button and reset the focus position by holding that for another 2-3 sec.
    • Open the black environmental control box to access the stage.

    • Turn on the light in the box, if needed.

    • Make sure that the objective is in the lowest position. If you are not sure, engage the escape button and reset the focus position by holding that for another 2-3 sec.

    • Move the condensor arm to the back.

    • Choose the frame insert from the tower - drawer Okolab frames

    • Place the insert into the stage and ensure that the red dot on the insert aligns with the red dot on the stage in the same corner.

    • Mount your sample and move the stage that the objective is below the sample.

  7. Choose the 4x Overview OC. Go Live Press the Autoscale button (Histogram with Play icon.) This will ensure that the Histogram will be scaled and you won't miss the sample even with a weak signal.
    • Choose the 4x Overview OC.

    • Go Live

    • Press the Autoscale button (Histogram with Play icon.) This will ensure that the Histogram will be scaled and you won't miss the sample even with a weak signal.

    • Alternatively, you can also select 4x objective and WF fluorescence channel for the overview if BF does not provide good contrast for sample localisation

    • Go up with the objective rotating the knob towards yourself.

    • The number of arrows indicates you the speed of movement. (3 arrows = fastest)

    • You can change the speed with this button

    • Start with 3 or 2 arrows and go up carefully, until you will hear gentle beeping with which microscope indicates you are near to the focal plane. Then change the speed to 1 arrow and find your focus.

  8. Once you focused on the slide/sample, localize the Sample Navigation tab Choose from the sample holder - if you cannot find the exact same one, choose from the most similar one.
    • Once you focused on the slide/sample, localize the Sample Navigation tab

    • Choose from the sample holder - if you cannot find the exact same one, choose from the most similar one.

    • Choose Current settings and Current Z position - this will use the last used OC, which you used for focusing in previous step

    • Click on Preview

    • Microscope will acquire a tilescan, which you can use later for the navigation in the sample

    • By clicking on Set you can update your focus position.

Finish Line

z mbstaff

Member since: 05/06/2019

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