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Updated: Sep 30, 2025

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
Statistical Considerations and Tools to Improve Histopathologic Protocols with Spectroscopic Imaging
Shachi Mittal1,2, Jonathan Kim3, Rohit Bhargava1,4,5
197472Department of Bioengineering and Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Abstract:
Advances in infrared (IR) spectroscopic imaging instrumentation and data science now present unique opportunities for large validation studies of the concept of histopathology using spectral data. In this study, we examine the discrimination potential of IR metrics for different histologic classes to estimate the sample size needed for designing validation studies to achieve a given statistical power and statistical significance. Next, we present an automated annotation transfer tool that can allow large-scale training/validation, overcoming the limitations of sparse ground truth data with current manual approaches by providing a tool to transfer pathologist annotations from stained images to IR images across diagnostic categories. Finally, the results of a combination of supervised and unsupervised analysis provide a scheme to identify diagnostic groups/patterns and isolating pure chemical pixels for each class to better train complex histopathological models. Together, these methods provide essential tools to take advantage of the emerging capabilities to record and utilize large spectroscopic imaging datasets.
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