Biology Methods and Protocols, cilt.10, sa.1, 2025 (ESCI, Scopus)
Histological embedding and staining techniques are essential for examining tissue and cellular morphology. This study compares two embedding methods - JB-4™, a glycol methacrylate-based resin, and conventional paraffin - to determine which method provides superior visualization of liver and long bone tissues under light microscopy. Liver tissues from both embedding protocols were stained using the Periodic Acid-Schiff method and silver impregnation method. JB-4 sections were also stained with acid fuchsin and toluidine blue, while paraffin sections were stained with hematoxylin and eosin staining. Contrary to the common assumption that JB-4 may interferes with certain staining protocols, acid fuchsin and toluidine blue yielded high-contrast, structurally detailed results in JB-4 sections. Both techniques preserved liver morphology. However, JB-4 demonstrated higher resolution and enhanced visualization of intracellular structures. JB4 also preservedglycogen more effectively. Cellular structures including nuclei, nucleoli, bile duct epithelial cells, and Kupffer cells, were observedmore distinctly in JB-4 preparations. Reticular fibers were similarly visualized with both embedding techniques. In contrast, paraffin embedding provided better preserved overall tissue architecture. Whilelong bone specimens, paraffin sections frequently displayed poorly defined structures, while JB-4 offered clearer visualization of chondrocyte lacunae, osteocyte nuclei, lamellar bone, and bone marrow cells. JB-4 and paraffin each offer distinct advantages depending on tissue type and histological objective. JB-4 appears to be compatible with a broader range of stains than was previously reported, which expands its utility in detailed tissue analysis. The selection of an embedding method should align with the morphological characteristics of the target tissue and the specific research goals.