Fixing Cells with Nature’s Nectar: Bee Honey versus Ethanol in Cytopathology

Background: Fixation is a crucial process in cytopathology that maintains cellular morphology for diagnosis. Ethanol is classic fixative for years. Honey, being antimicrobial, enzymatic, and acidic, is a possible eco-friendly alternative. Objective: The objective of the study was to assess the effectiveness of 20% and 50% concentrations of unprocessed honey as cytological fixatives relative to 95% ethanol, employing Hematoxylin and Eosin (H and E) and Papanicolaou (PAP) staining techniques. Materials and Methods: A comparative experimental study was done in the Department of Pathology on 240 cytological samples received over 4 months. Smears were divided along all three fixed, that is, 95% ethanol, 20% and 50% honey for 5 min, and were stained using H and E and PAP stains. The slides were evaluated using five cytomorphological criteria: clarity, uniformity, background, nuclear, and cytoplasmic details. The scores were added and ranked as poor (≤2), good (3–4), and excellent (5). Statistical comparison was carried out on R software v4.4.2 using McNemar’s test. P value was calculated. Results: Ethanol performed better than honey in clarity, nuclear and cytoplasmic staining for both stains ( P < 0.0001). Twenty percent honey provided greater clarity and clean background compared to 50% honey. For H and E staining, 20% honey registered excellent scores (88.3%), narrowly behind ethanol (87.5%). PAP-stained smears, ethanol provided the highest percentage of excellent-quality slides (85.41%), followed closely by 20% honey (74.58%). Conclusion: Ethanol remains a superior fixative in cytology. 20% honey can be used as a potential alternative fixative in cytopathology in limited-resource facilities. H and E had better staining quality than Pap in honey fixed smears. 50% honey should be avoided for poor-quality staining as compared to 20% honey.