<![CDATA[Formalin-fixed, paraffin-embedded (FFPE) tissues are widely used for histological and molecular analyses. Although fixative reuse can reduce costs and waste, its effects on DNA quality and downstream molecular applications remain a concern. This study investigated the effects of reusing 10% neutral buffered formalin (NBF) on H&E staining quality, DNA quality, and polymerase chain reaction (PCR) amplification. Four fixation conditions were evaluated: fresh NBF (0x; control) and NBF reused once (1x), twice (2x), or three times (3x), with six FFPE tissue sections assessed per condition. H&E staining quality was scored by a board-certified anatomic pathologist under blinded conditions. DNA concentration and purity were measured by spectrophotometry, DNA integrity was assessed by agarose gel electrophoresis, and PCR amplification of a 565-bp albumin gene fragment was evaluated. After three cycles of fixative reuse, 83% of tissue sections retained good H&E staining quality, with no statistically significant difference from the control (p>0.05). Good DNA integrity, however, was observed only in samples fixed with NBF reused up to twice. These findings suggest that NBF reuse should be limited to two cycles when FFPE tissues are intended for DNA analysis.]]>
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