Protocol for generation, quantification, and phenotyping of brain metastases in preclinical mouse models

The field of brain metastasis is rapidly expanding, yet no consensus exists on the most reliable quantification approach. We present a protocol for assessing metastatic burden in mice following intracarotid injection of tumor cells. We describe steps for surgical procedures, brain processing, cryo-sectioning, and slide preparation, followed by phenotypic characterization. We detail procedures for quantifying brain metastatic area using automated microscopy and semi-supervised image analysis. For complete details on the use and execution of this protocol, please refer to Rodriguez-Baena et al.¹.