Cell Proliferation Analysis
Imaging PCNA Staining for Cell Proliferation
Neonatal bone sections are common tissues to study chondrocyte proliferation. Immunohistochemical PCNA labeling in decalcified paraffin sections is fairly straightforward. Ideally, counter stains would b. adjusted to be relatively minimal. Too dark a counter-stain can sometimes be confused for strong PCNA signal.
BIOQUANT tracks the location of each measured cell to help ensure that no cell is counted twice. This can be done on a live image with a tracking stage, but it is simpler and more accurate to scan the section with either a scanning upgrade for your lab's microscope or with a digital pathology scanner in a core imaging facility.
Measure the Proliferation Zone
Define the Sampling Area
Define the contours of the proliferation zone using one of BIOQUANT's region of interest tools. The allows BIOQUANT to ensure that only chondrocytes within the proliferation zone are counted. This also records all the area data needed to normalize the number of proliferating cells by the size of the zone.
Automatic Region of Interest
Regardless of the magnification or location on the section, BIOQUANT recognizes the boundaries of the proliferation zone and automatically ensures that only cells within the zone are counted.
Measure Cells at High Magnification
Automatic Cell Detection
Using the DAB chromagen, BIOQUANT identifies PCNA+ chondrocytes. Size and shape filters exclude non-specific staining.
Using the counterstain, BIOQUANT identifies the other chondrocytes in the referent area.
Intelligent Filtering for Overlapping Fields of View
BIOQUANT maps the location of every counted cell in the field of view. This allows BIOQUANT to automatically filter out cells that have already been counted, even if they appear again in adjacent, overlapping fields of view.
At any stage, you may use the paintbrush or eraser tools to add, remove, or correct the shape of detected cells. Data are only recorded after you approve the software's cellular detection.
Proliferation Index and Other Data
BIOQUANT automatically computes:
- Total Sample Area
- Number PCNA+ Cells
- Number PCNA- Cells
- Total Cell Number
- PCNA+ Cell Density (#/mm2)
- PCNA+ Cell Optical Density (OD)
- Proliferation Index