After After
Before Before

10146 - CD57, Ovarian Cancer, TME

INTRODUCTION
Natural Killer (NK) cells are a subtype of lymphocytes. They hold anticancer potential through tumor cell lysing and their prognostic role is being investigated in many cancers, with a perspective of a therapeutic potential. The prognostic role of tumor infiltrating NK cells in ovarian cancer is being investigated in Department of Clinical Pathology, Vejle Sygehus.
This APP can be used to evaluate the number or density of CD57 positive nuclei.

KEYWORDS
CD57, NK cells, Natural Killer Cells, Ovarian Cancer, Cell Quantification, Image Analysis

METHODS
A NK positive cell is detected as a haematoxylin stained nucleus surrounded by DAB membrane stain. Initially, the protocol detects haematoxylin stained nuclei surrounded by brown stain using a poly blob filter on the H&E haematoxylin band and a median filter on the HDAB-DAB band. The DAB membrane stain is emphasized by a mean unsharp filter, thereby excluding blurry background stain. Then, positive nuclei are detected based on size, shape, and amount of surrounding DAB stain. Final NK cells are seen in Figure 2.

QUANTITATIVE OUTPUT VARIABLES
The output variables obtained from this protocol are:

  • # POS NK nuclei: Number of natural killer cells (CD57 positive nuclei)
  • ROI Area: Area of the outlined ROI
  • Positive Density: #POS NK nuclei / ROI Area

AUXILIARY APPs
No auxiliary APPs are available.

WORKFLOW
Step 1: Manually outline regions of interest (ROIs)
Step 2: Load and run the APP for the quantification of CD57 nuclei

STAINING PROTOCOL
Thermo Fisher Scientific, Invitrogen (Roche Ventana Benchmark platform).

ADDITIONAL INFORMATION
The APP utilizes the Visiopharm EngineTM and Viewer software modules, where EngineTM offers an execution platform to expand processing capability and speed of image analysis. The Viewer allows a fast review together with several types of image adjustment properties e.g. outlining of regions, annotations and direct measures of distance, curve length, radius, etc.
By adding the AuthorTM module the APP can be customized to fit other purposes. AuthorTM offers a comprehensive and dedicated set of tools for creating new fit-for-purpose analysis APPs, and no programming experience is required.

REFERENCES
USERS
This APP was developed for Dr. Jon Røikjær Henriksen, Department of Clinical Pathology, Vejle Sygehus.

RUO
Figure 1
Figure 1
Selected view of ovarian CD57 stained tissue.
Figure 2
Figure 2
Final classification of NK cells (red). Brown structures with no blue core are discarded as CD57 positive cells.