Problem
Diagnostic neuropathology traditionally relies on subjective interpretation of visual data obtained from a brightfield microscope. This results in high variability, inability to multiplex, and unsatisfactory reproducibility even among experts. These diagnostic problems may affect patient outcomes and confound clinical decision-making. Furthermore, standard histological processing of pathological specimens results in auto-fluorescence and other artifacts, which have nearly blocked the implementation of fluorescent microscopy in diagnostic pathology. Thus, generation of objective and quantitative methodologies would augment the toolbox available to neuropathologists, which would ultimately enhance clinical decision making.
Objective
To develop image analysis methods to quantitatively validate anti-PTBP1 antibody for use in diagnostic neuropathology.
Method
We propose a computerized image analysis method to validate anti-PTBP1 antibody. Images were obtained from standard neuropathological specimens stained with anti-PTBP1 antibody. First, the noise characteristics of the images were modeled and images are de-noised according to the noise model. Next, images are filtered with sigma-adaptive Gaussian filtering for local normalization, and cell nuclei are detected and segmented with a k-means based deterministic approach.
Result
Experiments on 29 data sets from three cases of brain tumor (recurrent glioma, primary resections of glioblastoma harboring the EGFRVIII mutation, pilocytic astrocytoma) and reactive gliosis show statistically significant differences between the number of positively stained nuclei in images stained with and without anti-PTBP1 antibody (p values, t-test, are 40×10−4, 33×10−4, 6×10−4 and 46×10−3, respectively).
Conclusion
The experimental of analysis of specimens from three different brain tumor groups and one reactive gliosis group indicate the feasibility of using anti-PTBP1 antibody in diagnostic neuropathology and computerized image analysis provides a systematic and quantitative approach to explore feasibility.