Guilherme R Ferreira, Livia Huang, Lori Strong, Geoffrey D Hugo, Buck E Rogers, Arash Darafsheh
Preclinical radiobiological research is essential for advancing the understanding of how different radiation delivery characteristics influence biological responses and for optimizing treatment strategies across diverse target sites. The precision, reproducibility, and translational relevance of these studies rely on the appropriate implementation of image-guided small animal irradiation platforms that accurately model clinical radiation therapy workflows. The current article describes the practical operation of two widely adopted image-guided preclinical radiation therapy systems used for high-precision small animal irradiation. Although both platforms are designed to deliver conformal radiation with image guidance, they differ in key engineering features, including X-ray tube specifications, source-to-axis distance (SAD), beam collimation, shielding mechanisms, mechanical geometry, and treatment planning software. These differences influence system setup, irradiation workflows, and treatment delivery. By outlining the methodological considerations and operational procedures for both platforms, this protocol provides investigators with practical guidance for designing, implementing, and reproducing translationally relevant preclinical radiation experiments.