Abstract
Bioluminescence imaging (BLI) is a highly sensitive tool for visualizing tumors, neoplastic development, metastatic spread, and response to therapy. Although BLI has engendered much excitement due to its apparent simplicity and ease of implementation, few rigorous studies have been presented to validate the measurements. Here, we characterize the nature of bioluminescence output from mice bearing subcutaneous luciferase-expressing tumors over a 4-week period. Following intraperitoneal or direct intratumoral administration of luciferin substrate, there was a highly dynamic kinetic profile of light emission. Although bioluminescence was subject to variability, strong correlations (r >.8, p <.001) between caliper measured tumor volumes and peak light signal, area under light signal curve and light emission at specific time points were determined. Moreover, the profile of tumor growth, as monitored with bioluminescence, closely resembled that for caliper measurements. The study shows that despite the dynamic and variable nature of bioluminescence, where appropriate experimental precautions are taken, single time point BLI may be useful for noninvasive, high-throughput, quantitative assessment of tumor burden.
Highlights
Bioluminescence imaging (BLI) is being rapidly adopted in cancer research [1]
Small changes in tumor volume occurred over this time, the variance in output profile and magnitude was independent of such changes
BLI is currently a 2-D representation of a 3-D tumor, and the signal intensity shows variations following administration of luciferin, we have found strong correlations between the measures of bioluminescent parameters and tumor volume
Summary
Bioluminescence imaging (BLI) is being rapidly adopted in cancer research [1]. The high sensitivity of bioluminescence technology permits the study of subtle biological events, those related to oncology such as early tumor development, metastases, residual disease, and tumor recurrence [2,3,4,5]. Tumor cells expressing luciferase from Photinus pyralis (firefly) are implanted subcutaneously in mice and subsequent growth is assessed over a period of days to weeks using BLI. As image acquisition times are short, such procedures can be completed in a highthroughput manner; little has been reported about the temporal nature of bioluminescence, the relationship between bioluminescence and tumor development, and the validity of bioluminescence as a highthroughput modality. Several investigations report a strong relationship between number of cells and bioluminescent intensity in culture [3,6,7]. Some investigations have examined increase in signal with tumor growth following implantation, but only few reports have provided independent measures of developing tumor volume [4,8]. Use of BLI as a quantitative tool remains to be thoroughly tested [1,10,11]
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