Protocol for noninvasive imaging of hematopoietic reconstitution in live mice using Akaluc bioluminescence

Here, we present a protocol for noninvasive, spatiotemporal visualization of hematopoietic reconstitution in live mice using Akaluc-based bioluminescence imaging (AkaBLI). We describe steps for Akaluc gene transduction into murine hematopoietic stem cells (HSCs), ex vivo expansion of hematopoietic stem and progenitor cells, and longitudinal imaging post transplantation. Representative results illustrate the spatiotemporal pattern of hematopoietic recovery in the spleen and bone marrow. This protocol enables quantitative in vivo tracking of hematopoietic reconstitution dynamics at whole-body resolution following lethal irradiation. For complete details on the use and execution of this protocol, please refer to Yogo et al.¹.