Measure Object Radial Distribution measures the radial distribution of intensities within each object.
Given an image with objects identified, this module measures the intensity distribution from each object's center to its boundary within a user-controlled number of bins, i.e. rings.
The distribution is measured from the center of the object, where the center is defined as the point farthest from any edge. The numbering is from 1 (innermost) to N (outermost), where N is the number of bins specified by the user. Alternatively, if primary objects exist within the object of interest (e.g. nuclei within cells), you can choose the center of the primary objects as the center from which to measure the radial distribution. This might be useful in cytoplasm-to-nucleus translocation experiments, for example. Note that the ring widths are normalized per-object, i.e., not necessarily a constant width across objects.
Available measurements
- FracAtD: Fraction of total stain in an object at a given radius.
- MeanFrac: Mean fractional intensity at a given radius; calculated as fraction of total intensity normalized by fraction of pixels at a given radius.
- RadialCV: Coefficient of variation of intensity within a ring, calculated over 8 slices.
See also MeasureObjectIntensity.
Settings:
Select an image to measure
Select the image that you want to measure the intensity from.
Select objects to measure
Select the objects that you want to measure the intensity from.
Object to use as center?
There are three ways to specify the center of the radial measurement:
- These objects: Use the centers of these objects for the
radial measurement.
- Centers of other objects: Use the centers of other objects
for the radial measurement.
- Edges of other objects: Measure distances from the
edge of the other object to each pixel outside of the
centering object. Do not include pixels within the centering
object in the radial measurement calculations.
For example, if measuring the radial distribution in a Cell
object, you can use the center of the Cell objects (
These objects)
or you can use previously identified Nuclei objects as
the centers (
Centers of other objects).
Select objects to use as centers
(Used only if "Centers of other objects" are selected for centers)
Select the object to use as the center, or select None to
use the input object centers (which is the same as selecting
These objects for the object centers).
Scale the bins?
Select Yes to divide the object radially into the number
of bins that you specify.
Select No to create the number of bins you specify based
on distance. For this option, the user will be
asked to specify a maximum distance so that each object will have the
same measurements (which might be zero for small objects) and so that
the measurements can be taken without knowing the maximum object radius
before the run starts.
Number of bins
Specify the number of bins that you want to use to measure
the distribution. Radial distribution is measured with respect to a series
of concentric rings starting from the object center (or
more generally, between contours at a normalized distance
from the object center). This number
specifies the number of rings into which the distribution is to
be divided. Additional ring counts can be specified
by clicking the Add another set of bins button.
Maximum radius
Specify the maximum radius for the unscaled bins. The unscaled binning
method creates the number of bins that you
specify and creates equally spaced bin boundaries up to the maximum
radius. Parts of the object that are beyond this radius will be
counted in an overflow bin. The radius is measured in pixels.