Phosphate-buffered Formalin Fixative Under Controlled Fixation Temperature Yielded Excellently Preserved Histomorphology

Abstract

Aim: Using mouse liver as experimental model, this study attempts to identify a formalin-based fixative and fixation temperature that jointly provides the best balance of preservation of tissue morphology. Methodology: Liver samples from fifty (50) albino mice aged between of 6 to 8 weeks consisting of both male and female was harvested following cervical dislocation and randomly distributed into control and experimental groups. Control samples were fixed in 10mL of 10% formalin at 25oC, 30oC, 35oC, 40oC, 45oC, 50oC, 55oC and 60oC respectively for 24 hours, while experimental samples were each fixed in equal volume of phosphate-buffered 10% formalin (pH 7.2, 7.4, 7.6 and 7.8) at the same temperature and time duration regimen and processed for general tissue morphology. Nuclear, cytoplasm and cell membrane morphology were assessed as evidence of the combined effectiveness of fixative and fixation temperature. Morphology was scored using a four-point grading scale with 1 being poor and 4 being excellent. Results: Nuclear, cytoplasm and cell membrane morphology were excellently preserved in tissue fixed with phosphate-buffered 10% formalin (pH 7.2) at 45oC. Tissue fixed with 10% formalin at 35oC exhibited excellent nuclear and cell membrane morphology, while excellent preservation of cell membrane morphology were observed in tissues fixed with 10% formalin at 40oC, phosphate-buffered 10% formalin (pH 7.4) at 55oC and 60oC, (pH 7.6) at 50oC and 55oC and (pH 7.8) at 55oC respectively. Furthermore, excellent preservation of nuclear morphology was observed in tissue fixed with phosphate-buffered 10% formalin (pH 7.8) at 60oC. Conclusion: Phosphate-buffered 10% formalin at a temperature of 45oC and pH 7.2 provide an excellent formalin-based fixative and fixation temperature that adequately preserves the microanatomy of tissue for histopathology examination.