Open topic with navigation

Preparing to trace

Before you start tracing a live image... Before you start tracing an imported image...
  1. Place a slide on the stage.
  2. Select objective and lens.
    1. Rotate the microscope's nose piece to select the appropriate objective.
    2. Select the matching lens from the Lens drop-down menu in the toolbar.
  3. Optional: If you don't see the tissue, select Acquisition>Live Image.
  4. Choose a location for the reference point.
    • Manual stage: Move the stage until the desired location is within the field of view.
    • Motorized stage: Use the joystick to move to the desired location. Using the Joystick
  5. Click to place the reference point.
  6. Verify that Neurolucida is registering movement along the Z-axis.
    1. Display the Z Meter by clicking the Z Meter icon .
    2. Focus up and down using the focus knob on the joystick (or the focus on the microscope if equipped with an internal Z motor).
  7. Click Trace>Manual Neuron Tracing.
  1. Open the image or image stack.
  2. If the image was acquired with an MBF system, go to step 3; if not, verify that the scaling is correct—This is critical!

    3. Correct scaling means that Neurolucida uses the same scaling (i.e., micron-to-pixel or micron-to-voxel ratio) as the scaling that was used when the image was acquired.

    1. If scaling is embedded in the image, Neurolucida applies the correct scaling.
      • If there is no software lens associated with this scaling, you are prompted to define a lens so that the image is displayed at its real/original size.
    2. If no embedded scaling is identified:
    3. Measure an artifact of a known length with Tools>Quick Measure Line to verify that Neurolucida is applying the correct scaling.
  3. Verify that Neurolucida is registering movement along the Z-axis.
    1. Display the Z Meter by clicking the Z Meter icon .
    2. Focus up and down using the mouse wheel or the Page Up/Page Down keys.
  4. Click Trace>Manual Neuron Tracing.

 

See Tracing Trees in Single Sections, Tracing Trees in Serial Sections, Tracing Contours, Serial Section Reconstruction

Neurolucida 11 | MBF Bioscience Support Center | Downloads