Staining and scanning of tissue samples for microscopic examination is fraught with undesirable color variations arising from differences in raw materials and manufacturing techniques of stain vendors, staining protocols of labs, and color responses of digital scanners. When comparing tissue samples, color normalization and stain separation of the tissue images can be helpful for both pathologists and software. Techniques that are used for natural images fail to utilize structural properties of stained tissue samples and produce undesirable color distortions. The stain concentration cannot be negative. Tissue samples are stained with only a few stains and most tissue regions are characterized by at most one effective stain. We model these physical phenomena that define the tissue structure by first decomposing images in an unsupervised manner into stain density maps that are sparse and non-negative. For a given image, we combine its stain density maps with stain color basis of a pathologist-preferred target image, thus altering only its color while preserving its structure described by the maps. Stain density correlation with ground truth and preference by pathologists were higher for images normalized using our method when compared to other alternatives. We also propose a computationally faster extension of this technique for large whole-slide images that selects an appropriate patch sample instead of using the entire image to compute the stain color basis.

Structure-Preserving Color Normalization and Sparse Stain Separation for Histological Images

The history of histology indicates that there have been significant changes in the techniques used for histological staining through chemical, molecular biology assays and immunological techniques, collectively referred to as histochemistry. Early histologists used the readily available chemicals to prepare tissues for microscopic studies; these laboratory chemicals were potassium dichromate, alcohol and the mercuric chloride to harden cellular tissues. Staining techniques used were carmine, silver nitrate, Giemsa, Trichrome Stains, Gram Stain and Hematoxylin among others. The purpose of this research was to assess past and current literature reviews, as well as case studies, with the aim of informing ways in which histological stains have been improved in the modern age. Results from the literature review has indicated that there has been an improvement in histopathology and histotechnology in stains used. There has been a rising need for efficient, accurate and less complex staining procedures. Many stain procedures are still in use today, and many others have been replaced with new immunostaining, molecular, non-culture and other advanced staining techniques. Some staining methods have been abandoned because the chemicals required have been medically proven to be toxic. The case studies indicated that in modern histology a combination of different stain techniques are used to enhance the effectiveness of the staining process. Currently, improved histological stains, have been modified and combined with other stains to improve their effectiveness.

/pdf/histological-stains-a-literature-review-and-case-study-3o0ca7hkh1.pdf

Histological Stains: A Literature Review and Case Study

Impacts of dietary exposure to different sized polystyrene microplastics alone and with sorbed benzo[a]pyrene on biomarkers and whole organism responses in mussels Mytilus galloprovincialis

Dietary chitosan nanoparticles enhance the growth, production performance, and immunity in Oreochromis niloticus

A Review for the Peripheral Nerve Interface Designer.

Managing the Laboratory Safety in the Laboratory Light Microscopy Fixation of Tissues The Gross Room/Surgical Cutup Tissue Processing and Microarray Microtomy: Paraffin and Frozen Plastic Embedding for Light Microscopy How Histological Stains Work The Hematoxylins and Eosin Connective and Mesenchymal Tissues and Stains Carbohydrates Pigments and Minerals Amyloid Microorganisms Bone Techniques in Neuropathology Immunohistochemical Techniques Immunofluorescent Techniques Immunohistochemistry Quality Control Molecular Pathology Transmission Electron Microscopy Quantitative Data from Microscopic Specimens Appendices

Bancroft's Theory and Practice of Histological Techniques

The hematoxylins and eosin

Connective and other mesenchymal tissues with their stains

Fixation of tissues

The gross room/surgical cut-up

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