Stereology Estimates

BIOQUANT Measurement’s cell module includes the Stereology Toolkit. Stereology Grids provide an established set of tools for the stereological estimation of cell number. Stereology templates are provided for the Optical Fractionator Project, Lung Stereology, and Cavalieri Stereology.

Optical Fractionator Project

CREATE A NEW PROJECT

Use the Optical Fractionator Project Manager to clearly and easily define a new project using Stereological methods.

TRACE THE SAMPLING AREA

Either on a montage or at low magnification, the sampling area is traced. The grid is automatically bound to the sampling area and sampling locations within the sampling area are automatically marked.

USE THE GRID WIZARD

Specific grid dimensions are memorized per project.

SET UP THE DISECTOR

At high magnification the counting frame is drawn on the live image making it easy to count neurons within the counting space. The rotator or nucleator probe can also be used at this point to estimate the mean cell size and population of cells.

AT EACH DISECTOR LOCATION, COUNT CELLS

Count cells at each disector location by clicking on each valid cell nuclei you see. Cell Number Estimates are made using the Optical Fractionator, Optical Disector, or Physical Disector. Total Length or Surface Area Estimates are made with cycloid curves or weighted line patterns on vertical or IUR sections. Reference Volume Estimates are made using the Cavalieri method.

RECORD MEAN CELL VOLUME ESTIMATES

Mean Cell Volume Estimates are made using the Rotator or Nucleator probes on vertical or IUR sections.

ERROR ESTIMATION

Error Estimation is built-in and revised as needed for optical fractionator estimates of cell number. All raw data can be exported to allow for custom error estimation methods.

Lung Stereology

LUNG TISSUE MORPHOLOGY

Using a fractionator sampling scheme, sampling locations are chosen over the tissue, allowing a smaller sample size to represent the whole lung tissue volume.

Within individual dissectors, a pattern of points and orientation weighted lights is randomized and superimposed on the tissue.

Intersections between the grid points within the dissector and the underlying tissue perform a cavaleri estimate of tissue and airspace volume.

Intersections between the lines and the alveolar surface perform as well as the number of end points and triple points provide an estimate of alveolar surface length.

COMPUTED DATA

Total Volume
Tissue Volume
Air Volume
Triple Point Count
End Point Count
Inter-alveolar Septal Length
Alveolar Surface

Cavalieri Stereology

VOLUME RATIO ESTIMATES

In serially sectioned tissue, the relative volumes of different types of tissue are quickly estimated using a regularly spaced and randomly positioned grid.

The grid is projected onto the sample and the operator looks for grid intersections that fall on the type of tissue to be measured.

This is ideal for tissues where is it difficult to use a protein-specific stain to indicate the tissue to be measured. Lacking a specific stain, a human being has to recognize the tissue and decided if it falls under the grid intersections.

The total number of points in the grid estimate the total area of the tissue. The number of points over the target regions indicate the target area.

CALCULATED DATA

Total Number of Grid Intersections
Estimated Total Tissue Volume
Number of Specific Grid Intersections
Estimated Specific Tissue Volume
Estimated Volume Ratio Estimate

Bone Referent Data Stereology

Stereology is the use of statistical sampling to estimate the morphology of structures. It is particularly helpful when limited stain contrast makes automated analysis difficult.

Calibrated Image Montage

Calibrated Image Montage puts a single image of the entire sampling area (e.g. tissue volume) in the Image window, making it simple to accurately draw the sampling area boundary.

Grid Wizard

The Grid Wizard automatically generates sampling grids bounded by the Tissue Volume. Grids automatically re-size with changes in zoom level on the digital scan, or magnification if using live imaging.

MERZ Estimators

Count intersections between the bone surface and the grid lines to estimate bone surface in trabecular bone.
Count intersections between the bone and the points within the grid to estimate bone volume