Flexible Measurement of Bioluminescent Reporters Using an Automated Longitudinal Luciferase Imaging Gas- and Temperature-optimized Recorder (ALLIGATOR)

摘要: Luciferase-based reporters of cellular gene expression are in widespread use for both longitudinal and end-point assays biological activity. In circadian rhythms research, example, clock fusions with firefly luciferase give rise to robust bioluminescence that persist over many days. Technical limitations associated photomultiplier tubes (PMT) or conventional microscopy-based methods quantification have typically demanded cells tissues be maintained under quite non-physiological conditions during recording, a trade-off between sensitivity throughput. Here, we report refinement prior allows long-term imaging high throughput which supports broad range culture conditions, including variable gas humidity control, accepts different tissue plates dishes. This automated gas- temperature-optimized recorder (ALLIGATOR) also the observation spatial variations across cell monolayer tissue, cannot readily observed by traditional methods. We highlight how ALLIGATOR provides vastly increased flexibility detection activity when compared existing