pH and Osmolarity: The pH and osmolarity of the fixative should be close to physiological levels to prevent cellular damage. Temperature: Higher temperatures can accelerate fixation but may also cause tissue distortion. Fixative Concentration: The concentration of the fixative must be optimized to ensure adequate penetration and fixation without causing excessive hardening or shrinkage. Fixation Time: Adequate time is necessary for complete fixation, but over-fixation can lead to tissue brittleness. Tissue Size: Smaller tissue samples fix more quickly and thoroughly than larger ones.
