Honey as a Cytological Fixative: A Comparative Study With 95% Alcohol

Abstract

BackgroundNinety-five percent (95%) ethyl alcohol (ethanol) has been used as a standard cytological fixative but it is expensive, difficult to procure, and has addictive properties. Alternate substitutes like methanol, which give similar results to ethanol, have toxic potential. Honey, a known preservative, is an eco-friendly fixative and is of great value when ethanol is unavailable and economizing on cost is necessary. The present study was done to assess and compare the fixation property and cytomorphological features of smears fixed in 20% honey in comparison to 95% ethyl alcohol and to determine whether the former can be used as an alternative cytological fixative in routine practice. Material and methodsThe present prospective study was done in the cytology section of the Department of Pathology for one and a half years on 300 cytological samples comprising 100 samples each of various body fluids (peritoneal, pleural, bronchoalveolar lavage, and urine), cervical smears, and fine-needle aspiration samples. The smears from all the 300 cytological samples were fixed separately in 95% ethanol and 20% unprocessed natural honey for a minimum of 15 minutes and were then stained with Papanicolaou (Pap) stain. The cytomorphological parameters of both the smears were compared based on set criteria. Relevant statistical analysis was done using the student t-test, chi-square test, and test of agreement (kappa statistics). ResultsA comparable and good-quality staining pattern, preservation of morphology, and crisp nuclear and cytoplasmic staining were observed between the two fixatives for all three types of samples with a strong agreement between them (kappa value varying between 0.896 and 0.942) and a p-value of <0.05. ConclusionNatural honey is a readily available and non-toxic alternative to ethanol as a cytological fixative and can be used in routine practices, especially in resource-constrained settings.

Cadaver preservative properties of a solution composed of honey, ethyl alcohol, liquid paraffin, distilled water and citric acid: Experiments on rabbit cadavers

“…Sections from cadavers fixed with Thiel's solution, saturated saline solutions and SEFS were reported to have sufficient histomorphological quality (Hayashi et al, 2014(Hayashi et al, , 2016Lombardero et al, 2017;Turan et al, 2017). The solution exhibited satisfactory cytoplasmic and nuclear staining, retained cell morphology and excellent visibility in oral smear (Priyadarshi et al, 2022). Moreover, we investigated the potential preventive effects of a solution composed of honey, citric acid, ethyl alcohol and lavender essence on the sheep's heart in the pilot study.…”

“…Another study reported natural honey serves as a conveniently accessible and non‐toxic substitute for ethanol when used as a cytological fixative. The solution exhibited satisfactory cytoplasmic and nuclear staining, retained cell morphology and excellent visibility in oral smear (Priyadarshi et al., 2022). Moreover, we investigated the potential preventive effects of a solution composed of honey, citric acid, ethyl alcohol and lavender essence on the sheep's heart in the pilot study.…”

Comparison between 70% ethyl alcohol and 10% formalin as fixative mediums in surgical cooperation campaigns: a pilot study

“…In addition, they offer better DNA, RNA, and protein integrity (they are coagulant fixatives) and more stable specimen preservation -if stored for prolonged periods-when compared to formalin [1,7,10,12]. However, ABF are not free of disadvantages: they are considered highly flammable products, they evaporate quickly, and sometimes they can associate shrinkage of the fixed specimen, which in turn can lead to variability in the stains performed [5,6,9,10,13,14].…”

“…Of these natural alternatives, bee honey is the one that has shown better results [4]. 10 to 20% dilutions of bee honey have contrasted antibacterial, acidic, and dehydrating properties, and they have demonstrated a similar fixation profile to formalin and alcoholic fixatives, even for nuclear structures [1,3,4,11,[14][15][16].…”