The Effect of Fixation Time Variation on Erythrocyte Morphology in Pheriperal Blood Semar (Pbs) Using Giemsa Staining

Salmon is a highly nutritious food source that is widely consumed, but it is also vulnerable to contamination by Background: Peripheral Blood Smear (PBS) is an essential hematological examination to evaluate blood cell morphology. Fixation prior to staining plays a vital role in preserving erythrocyte structure. The variation in fixation time is suspected to influence erythrocyte morphology. Objective: To determine the effect of fixation time variation (1 minute, 2 minutes, and 4 minutes) on erythrocyte morphology based on shape, size, and color using Giemsa staining. Methods: This study is a laboratory experimental design using 30 PBS samples fixed with absolute methanol for 1, 2, and 4 minutes. The slides were stained using the Giemsa method and observed microscopically for erythrocyte morphology. Data were analyzed using the Kruskal-Wallis test and Mann- Whitney post hoc test with Bonferroni correction. Results: There was a significant difference in erythrocyte morphology in terms of shape, size, and color across fixation times (p < 0.05). The 2-minute fixation time produced the best morphology, with 100% good shape, normocytic size, and normochromic color. Meanwhile, at 4 minutes, there was an increase in poor crenation, microcytic and macrocytic size, and hypochromic and hyperchromic staining. Conclusion: Fixation time variation significantly affects erythrocyte morphology. Two minutes of fixation is the most optimal duration to maintain good erythrocyte morphology with Giemsa staining.