# Propolis as a Potential Novel Histological Tissue Fixative: A Preliminary Analysis
## 3 Results
### Comparison of Fixation Ability of Different Solutions throughout Various Microscopic Structures  
There was no significant difference in the mean score of almost all the microscopic structures individually. However, tissue samples placed in 6.6% propolis (Group P) showed better quality in superficial one-third of the epithelial lining with a significantly higher mean score compared to tissue placed in 0.9% saline (\(p\) = 0.03) (Table 6a-c). \begin{table}
\begin{tabular}{c c c c c c c} \hline \hline  & \multicolumn{6}{c}{**(a)**} \\ \hline \multirow{2}{*}{**Solution**} & \multicolumn{6}{c}{**Epithelium**} \\ \cline{2-7}  & \multicolumn{2}{c}{**Superficial 1/3 rd**} & \multicolumn{2}{c}{**Middle 1/3 rd**} & \multicolumn{2}{c}{**Lower 1/3 rd**} \\ \cline{2-7}  & **Mean** & **SD §** & **Mean** & **SD** & **Mean** & **SD** \\ \hline
0.9\% Saline & 2.58 & 0.44 & 2.83 & 0.30 & 3.42 & 0.10 \\ \hline
10\% Formalin & 3.12 & 0.39 & 3.21 & 0.29 & 3.41 & 0.22 \\ \hline
6.6\% Propolis & 3.37 & 0.16 & 3.21 & 0.21 & 3.41 & 0.22 \\ \hline
6.6\% Propolis + 10\% Formalin & 3.16 & 0.36 & 3.21 & 0.16 & 3.50 & 0.36 \\ \hline \multicolumn{7}{c}{Pairwise comparisons by [PERSON]’s multiple post hoc procedures} \\ \hline
0.9\% Saline vs. 10\% Formalin & \(p\) = 0.2342 & \(p\) = 0.4502 & \(p\) = 1.0000 \\ \hline
0.9\% Saline vs. 6.6\% Propolis & \(p\) = 0.0388 * & \(p\) = 0.4502 & \(p\) = 1.0000 \\ \hline
0.9\% Saline vs. 6.6\% Propolis + 10\% Formalin & \(p\) = 0.1779 & \(p\) = 0.4502 & \(p\) = 0.9859 \\ \hline
10\% Formalin vs. 6.6\% Propolis & \(p\) = 0.8374 & \(p\) = 1.0000 & \(p\) = 1.0000 \\ \hline
10\% Formalin vs. 6.6\% Propolis + 10\% Formalin & \(p\) = 0.9998 & \(p\) = 1.0000 & \(p\) = 0.9842 \\ \hline
10\% Propolis vs. 6.6\% Propolis + 10\% Formalin & \(p\) = 0.9080 & \(p\) = 1.0000 & \(p\) = 0.9842 \\ \hline \hline \end{tabular}
\end{table} 
Table 6: (a) Comparison of fixative ability of different solutions in different microscopic structures (epithelium). (b) Comparison of fixative ability of different solutions in different microscopic structures (basement membrane, nerve bundles, and fibrous tissue). (c) Comparison of fixative ability of different solutions in different microscopic structures (blood vessels, skeletal muscles, and adipose tissue). Figure 7: A graph showing within-group comparison at different time points. At the microscopic level, tissues placed in 0.9% saline showed significant cellular swelling (cytoplasmic vacuoization), shrinkage of the nucleus (pyknotic), indistinct cell borders, and poor staining quality. These features may indicate tissue autolysis (Figure 8A). On the other hand, other fixative agents could fix tissue samples over different time points (Figure 8B-D). Most of the tissue samples placed in 6.6% propolisis showed better staining quality, cellular outline, and nuclear and cytoplasmic details than those in 10% formalin. However, homogenization of connective tissue was observed in some tissue samples placed in 6.6% propolisis for 48 h. The tissue sections of Group P + F had adequate overall morphology and nuclear, cytoplasmic details, and staining quality. Hence, all solutions, except 0.9% saline, were able to fix the tissue throughout different time points, with 6.6% propolisis giving the best results.