Lane Background Method

The Image Studio Lane background method works by drawing a linear connection between areas of minimum brightness between shapes in a lane and subtracting the background noise that falls beneath the lines.

The Lane background procedure outlined below requires more extensive explanation than the other background methods.

  1. Lane Center Found for Analysis

    The Image Studio Lane background subtraction procedure analyzes the middle 75% of a lane to compensate for variable shape width throughout the lane. The program uses the center lane line to establish the correct area to include in analysis. The lane line is first centered approximately by the user and then optimally positioned by the software during band-finding (the software will find the exact middle of the lane, do not worry about getting the line exactly positioned).

  2. Average Pixel Intensity Computed

    Next, a lane profile is created (see image below)(see . Representation of the lane profile calculation in Image Studio™ Software. Using the Profiles Tab to Inspect a Lane Profile for how to display a lane profile.). The lane profile is the set of average pixel intensities across every row of pixels in the center 75% of the lane.

    Image Studio lane profile blot to graph comparison

    3. . Representation of the lane profile calculation in Image Studio™ Software. Using the Profiles Tab to Inspect a Lane Profile for how to display a lane profile.

  3. A Background Curve Is Calculated

    Background is then established by drawing a linear connection between local minima in the Intensity vs. Location graph, as long as the slope of the line is below a threshold value. Limiting the slope prevents the possibility of drawing lines through a significant portion of a signal peak. Minima that would create a slope that is too steep are skipped, and the next point that would create a line with a slope below the threshold value is used.

    Image Studio Software will not seek local minima in shapes that have been added manually. Manually added shapes are skipped, because they have been designated as important by the user.

    4. Image Studio lane background calculation example
    An example Lane background calculation for shape B02 is shown using fictitious numbers.

  4. Background Subtracted

    The background curve value at the top and bottom of a shape are averaged to find the background value. The background value is subtracted from the Total of the shape to find the Signal for the shape.

    Image Studio lane background calculation example