Abstract 7427: Automated, real-time acquisition and quantification of peak, plateau-phase in vivo bioluminescent data
Authors: Van Praagh Andrew , Nelson Bo , Ballieu Paul , Smith Melanie , Rule Mike 
Published: March 2024

Abstract
It is widely recognized that plateau-phase bioluminescent (BL) kinetic curve data should be used when reporting on BLI results from substrate-injection BLI model systems. The basis for this understanding is that plateau-phase BL data is uniquely suited in several ways: (i) it will optimize the detection of target, luciferase-expressing cells (especially when they are low in copy number or deep within the animal model), (ii) it will optimize BLI data reproducibility (with there being no change in signal values across the plateau phase), and (iii) it will also provide BLI data values that correlate consistently with the number of viable target cells in a given BLI model. Given this critical value of plateau-phase BL data, there is, nevertheless, a simple practical issue to be addressed: The overall manual process of generating BL kinetic curves, and then identifying and quantifying plateau-phase data is not insignificant—it takes time!In this poster, we review the performance of “Kinetics,” a new feature in the Aura software platform from Spectral Instruments Imaging, LLC. Kinetics is designed to collect, present, and analyze BL kinetic curve data for up to 10 mice, in a completely automated and real-time fashion. Data outputs include a live graph that simultaneously presents individual mouse, whole body ROI (Total photon/sec) values vs. time post-luciferase injection, and analogous mean BL values per mouse group. Here in, we present an evaluation of the performance of Kinetics under various testing conditions. We initially evaluated Kinetics’ ability to acquire, present and analyze the BL kinetic curves of 5 phantoms. The photon output rates of these phantoms (Total photons/sec) were programmed to mimic the rise, plateau, and fall sequence typically seen BLI studies. In an analogous fashion, we then tested Kinetics’ ability to acquire, present, and analyze the BL kinetic curves of 5 to 10 live mice from several oncology models. Furthermore, we used Kinetics in oncology efficacy studies to monitor for expected changes in BL kinetic curve shapes (i.e., for changes in the onset times and durations of the rise, plateau and fall phases in BL kinetic curves) between different treatment groups (e.g., between positive controls groups and one or more treatment groups). In the same studies, we also used Kinetics to check for changes in BL kinetic curve shapes that might occur for a given treatment group across several time points, after pathogen challenge. Results from this set of experiments consistently and clearly illustrated the capability of Kinetics to acquire, present and analyze individual and mean BL kinetic curve data for up to 10 mice at a time, in an automated and real-time fashion. We believe that Kinetics from Spectral Instruments Imaging, LLC, has the potential to revolutionize the ease with which plateau-phase, BL kinetic curve data can be collected, presented, and used in a wide range of preclinical BLI studies.
Citation Format: Andrew Van Praagh, Bo Nelson, Paul Ballieu, Melanie Smith, Mike Rule. Automated, real-time acquisition and quantification of peak, plateau-phase in vivo bioluminescent data [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7427.