8. View the sampling results

• Parameters: File information and parameters associated with the selected probe runs.
• Marker X: Population estimate(s) and total number of markers. Typically, each marker type represents one object type.
• Counts by Site: Raw data for each counting site visited in each of the runs.
• CE Gundersen/CE Scheaffer/CE Cruz-Orive/CE Schmitz-Hof: See Coefficients of Error.
• Z Depth Histogram: Z depth location for markers placed in each section.

• Planimetry: The area within each traced contour and the volume of the region of interest based on the total area of the contours and the virtual section height.

  Note that planimetry is biased; for an unbiased estimate of area/volume, use the Cavalieri Estimator.

The following results are displayed when Parameters category is selected in the left hand panel of the Sampling Results window:

• Data File Name: File name associated with this data set, if the data was already saved.
• Date and Time: When the probe was completed.
• Region: Name of the contour type that defines the region of interest. If this is a composite of several runs, displays the contour name used for the first run is shown.
• Number of Sampling Sites: Number of sampling sites visited on all selected sections.
• Disector Height: Z-axis height of each disector.

• Disector Volume: Area of the disector based on the counting frame dimensions and the disector height.

• Contour Shape Factor: (Average of all contours?) Shape factor provides information about the complexity of a contour and is calculated by dividing the perimeter of a contour by the square root of its area. Perimeter and area can be found in the contour measurement window.

• Counting Frame Width (µm): X-axis width of each counting frame.
• Counting Frame Height (µm): Y-axis height of each counting frame.
• Counting Frame Area (µm²): Area of a single counting frame.
• Sampling Grid Width (µm): Distance between counting frames (sampling sites) along the X-axis.
• Sampling Grid Height (µm): Distance between counting frames (sampling sites) along the Y-axis.
• Sampling Grid Area (µm²): Area of each grid step (Sampling Grid Width x Sampling Grid Height).
• Virtual Section Thickness (µm): Value used for section thickness across all sections that were sampled.
• Section Evaluation Interval: The interval of the tissue sections used for counting. Since Image Volume Fractionator uses virtual sections that are stacked directly on top of each other (rather than physically cut sections which can be separated out into interval sets), the section evaluation interval always equals 1.

Print Displayed Results: Prints the currently displayed results category for all selected probe runs.

Print All Results: Prints results of all selected probe runs.

Copy Displayed Results to Clipboard: Copies the currently displayed results category for all selected probe runs to the Windows Clipboard.

Copy All Results to Clipboard: Copies results of all selected probe runs to the Windows Clipboard.

Edit Shape Factor: Displays the Shape Factor dialog box (see Shape Factor). To obtain as accurate an estimate of the Coefficient of Error as possible, use the slider to edit the Shape Factor which describes the shape of the region of interest.

Edit Mounted Thickness: Use to adjust the mounted section thickness.

Equations: Displays the equations used for the probe runs.

Export to Excel: Exports results of all selected probe runs directly to Excel (2003 or later).

Close: Closes the Sampling Results window.

• Summary: Estimates and Coefficients of Error (CE) for each individual marker.
• Parameters: File information and parameters associated with the selected probe runs. This information should be added to the Methods section of a publication to enable other researchers to test the reproducibility of the results.
• Counts by Site: Lists the measured thickness and number of markers at each site. This information can be used to calculate your own CE or to compare thicknesses/number of objects within a section or between sections.
• Coefficient of Error: Lists all Coefficients of Error.
• CE Variance Details: Information related to the calculation of the Gundersen and Shaeffer CEs.
• Section Details: Lists marker counts by section.
• All Markers Z Histogram and Individual Markers Z Histogram: Distribution of objects within the tissue.

  Ideally, with no sectioning artifact from the microtome blade, there is an equal number of markers placed at each “bin” from the top of the site.

  In practice, there are fewer markers at the bottom/top of each site; set your guard zones so that they cover these regions.

  A spike of cells at the top of the histogram could be caused by focusing through the tissue or counting cell bottoms at the top of the tissue as though they were cell tops (assuming that cell tops are the unique point you're trying to count).

  A lower number of objects marked in the middle of the histogram is probably due to an incomplete staining penetration.

• Z Depth Details: Lists the Z values for each marker. These values are used to generate the Z depth histograms.
• Smoothness : Distribution of markers within your region of interest.
• Planimetry : Area of each section and volume calculated from this area. Note that this information is biased; for an unbiased estimate of area/volume, use the Cavalieri Estimator.
• Z Order : Z value of the sections and the actual Z at which the contours were drawn in each section.
• Raw Report : Number of markers and tissue thickness per site.