1. Inadequate Fixative Volume: Insufficient volume of fixative relative to the tissue size can result in poor penetration. 2. Large Tissue Size: Thick or large tissue samples can impede fixative penetration, leading to incomplete fixation. 3. Insufficient Fixation Time: Rushing the fixation process or not allowing sufficient time can leave deeper tissue layers unfixed. 4. Inappropriate Fixative: Using a fixative that is not suitable for the specific tissue type or study purpose can lead to suboptimal results. 5. Poor Fixative Quality: Degraded or contaminated fixatives can also cause incomplete fixation.
