Effects of Perfusion Volume and Delivery Technique on Rat Brain Histological Quality: A Comparison of Transcardial Perfusion Protocols

Makkiyah, Feda Anisah; Wahyudi, Dhyani Paramita; Amalia, Muttia; Pradana, Dhigna Luthfiyani Citra; Rahmi, Eldiza Puji; Buana, Gherriandi Raja

doi:10.33899/ijvs.2026.169622.4689

	Effects of Perfusion Volume and Delivery Technique on Rat Brain Histological Quality: A Comparison of Transcardial Perfusion Protocols
Iraqi Journal of Veterinary Sciences
Volume 40, Issue 3, July 2026, Pages 589-599 PDF (1.14 M)
Document Type: Research Paper
DOI: 10.33899/ijvs.2026.169622.4689
Authors
Feda Anisah Makkiyah^* ¹; Dhyani Paramita Wahyudi²; Muttia Amalia³; Dhigna Luthfiyani Citra Pradana²; Eldiza Puji Rahmi²; Gherriandi Raja Buana²
¹Jalan Rs Fatmawati 1
²Faculty of Medicine, Universitas Pembangunan Nasional “Veteran” Jakarta, South Jakarta, Indonesia
³Department of Clinical Pathology, Faculty of Medicine, Universitas Pembangunan Nasional “Veteran” Jakarta, South Jakarta, Indonesia
Abstract
Transcardial perfusion is widely used to preserve rodent brain tissue. However, standardized protocols regarding parameters such as fixative volume and delivery technique that ensure optimal histological quality have not yet been established. This study aims to evaluate the effects of fixative volume and delivery technique on the histological quality of rat brain tissue. Sixteen male Sprague-Dawley rats were allocated to four groups: manual-200 mL, manual-650 mL, pump-200 mL, and pump-650 mL. Transcardial perfusion was performed using specific methods for each group. Brains were processed with hematoxylin-eosin staining and photographed. Quantitative assessment of nuclear & cytoplasmic staining intensity was performed using Fiji. Histological artifacts (e.g., blood pooling) were scored. Group differences were analyzed using IBM SPSS Statistics. Nuclear staining intensity was highest in the manual-200 mL group (103.31 ± 19.29) and lowest in the manual-650 mL group (156.10 ± 14.94). Welch ANOVA confirmed a statistical difference (p<0.001) between the manual-200 mL and manual-650 mL groups and between the pump-200 mL and manual-650 mL groups. Cytoplasmic staining intensity showed the same trend. Tukey HSD indicates significance (p<0.001) between the same groups, as well as manual-200 mL and pump-650 mL. Artifact counts were descriptively higher in the pump groups but showed no significant association (p = 0.362). A lower volume of 200 mL and manual administration resulted in better histological quality during transcardial perfusion.
Keywords
brain model; fixation; hematoxylin-eosin; Sprague-Dawley; transcardial perfusion

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