Continuous and real-time in vivo autobioluminescent imaging in a mouse model

In vivo small animal bioluminescent imaging has become an indispensable technique for interrogating the localization, health, and functionality of implanted cells within the complex environment of a living organism. However, this task can be daunting for even the most experienced researchers because it requires multiple animal handling steps and produces differential output signal characteristics in response to a number of experimental design variables. The recent emergence of autobioluminescent cells, which autonomously and continuously produce bioluminescent output signals without external stimulation, has the potential to simplify this process, reduce variability by removing human-induced error, and improve animal welfare by reducing the number of required needlesticks per procedure. This protocol details the implantation and imaging of autobioluminescent cells within a mouse model to demonstrate how cells implanted from a single injection can be imaged repeatedly across any post-implantation timescale without the need for further human–animal interaction or signal activation steps. This approach provides a facile means to continuously monitor implanted cellular output signals in real-time for extended time periods.