A simple two-dye basic stain facilitating recognition of mitosis in plastic embedded tissue sections

Area of Science:

• Oral histology
• Cell biology
• Histopathology

Background:

• Accurate identification of mitotic figures in oral epithelium is crucial for understanding tissue proliferation and pathology.
• Conventional staining methods for semithin plastic sections often lack the contrast needed for clear visualization of cellular components in densely packed oral epithelium.

Purpose of the Study:

• To develop and evaluate a novel differential staining technique for semithin sections of buccal and palatal mucosa.
• To improve the recognition of mitoses and distinct cellular structures within the oral epithelium.

Main Methods:

• Semithin sections of oral mucosa were fixed with glutaraldehyde and osmium tetroxide, then embedded in Durcupan resin.
• A two-step staining procedure involved malachite green followed by crystal violet.
• Staining conditions included specific concentrations, temperatures, and incubation times for both stains.

Main Results:

• The staining method provided vivid purple contrast for nuclear envelopes and chromatin against a green cytoplasm.
• Chromosomes of dividing cells appeared bluish violet, and nucleoli showed varying greenish-blue hues.
• Distinct differential staining facilitated clear recognition of mitotic figures, even in crowded proliferative compartments.

Conclusions:

• This malachite green and crystal violet staining technique offers superior differentiation of cellular components in oral epithelium.
• It effectively aids in the identification of mitoses, addressing limitations of standard stains for plastic-embedded sections.
• The method holds potential for enhanced diagnostic accuracy in oral pathology and research.