Image Volume Fractionator workflow >

8. View the sampling results

Procedure

View the results using one of the following methods:

  • Click a set in the Probe Runs list to select it and click the View Results button. The Sampling results window is displayed.
  • Click Display Probe Run List (use after running more than one probe if you want results from multiple probe runs). The Previous Stereological Runs window is displayed.
    1. From the list, highlight the probe runs of interest.
    2. Click View Results. The Sampling results window is displayed.

See Display Probe Run List for a general description of the parameters displayed in the Sampling Results window and the Excel worksheet (if you elected to export the results to Excel).

Interpretation

Stereo Investigator Cleared Tissue Edition provides four estimates. The results vary based on the measurement used for the mounted (or post-processing) section thickness. The mounted section thickness value is divided into the counting frame thickness (or disector height) to calculate the height sampling fraction (hsf). We recommend that you report the estimate that best reflects the histological properties of the region of interest.

Estimated population using user-defined section thickness

Calculated using a single value entered manually for the post-processed or “mounted” section thickness.

Because this estimate is generated with only one value for the section thickness, local variations in section thickness are not accounted for. As a result, this estimate should be considered the least accurate of the 4 available estimates. But if there is no section thickness variation (e.g., embedding protocols such as plastic embedding), reporting the Estimated population using user-defined section thickness is acceptable.

This value is typically entered manually, or calculated, in the 5. Measure mounted thickness step of the workflow under the Manually enter the average mounted thickness method.

  • If you don't enter a value for Manually enter the average mounted thickness, the estimate equals zero.
  • To change the thickness value after the counting procedure, click the Edit Mounted Thickness button in the Sampling results window.

Estimated population using mean section thickness

If you didn't measure the thickness of sections while counting, this estimate is not calculated.

Calculated using the section thickness measurements recorded while counting.

These measurements are recorded in the 5. Measure mounted thickness step of the workflow, after selecting Measure the mounted thickness while counting method.

The number of measurements used to calculate this estimate is based on the interval you entered (e.g., if you entered 2, you are prompted to set the top and bottom of the section at every other counting site). The measured thickness values from these sites are averaged to produce a mean measured thickness value used for the height sampling fraction calculation.

Because this estimate is generated from the mean of all obtained section thickness measurements, it is considered to be the most accurate estimate of the region of interest when measurements are not performed at every site.

You may also choose to use this value for low frequency events (e.g., BrdU+ neurons) with many counting sites containing zero objects when you want to generate an accurate average measured section thickness measurement for the hsf using a systematic interval for site measurement.

Estimated population using mean section thickness (only using sites with counts)

If you didn't measure the thickness of sections while counting, this estimate is not calculated.

This estimate is a variation of Estimated population using mean section thickness.

Calculated using only the section thickness measurements made at counting sites that contain marked objects (in other words, section thickness measurements from counting sites with NO counted objects aren't included in the calculated average).

These measurements are recorded in Step 5 of the Optical Fractionator workflow, after selecting Measure the mounted thickness while counting method.

In many cases, this estimate will be nearly identical to Estimated population using mean section thickness.

You may choose to use this value when :

  • You chose to ignore measuring the section thickness where there were no objects.
  • You made errors in section thickness measurement that were not corrected when there were no objects to be marked.

Estimated population using number weighted section thickness

Report this estimate when thickness was measured at every sampling site and when the section thickness varies dramatically across the sections that include the region of interest.

Calculated using only the section thickness measurements from counting sites that contain markers. These measured thickness values are then weighted by the number of objects associated with them to produce a weighted average.

The number weighted mean section thickness is reported in the Parameters section and is used to calculate the height sampling fraction.

All estimates calculated from measurements obtained while counting (i.e., all but Estimated population using user-defined section thickness) should return similar results if the measurements were taken correctly.

Understanding the results in the Sampling Results window

 

 

See Display Probe Run List