THE SPATIAL DISTRIBUTION OF LIGHT EMISSION FROM LIQUID PHASE BIO- AND CHEMILUMINESCENCE: VARIATIONS WITH CONTAINER TYPES, TURBIDITY AND CONTAINER FROSTING

Abstract— The spatial distribution of emitted light from liquid phase bioluminescence and chemiluminescent sources in three common types of containers has been evaluated. These sources, while theoretically isotropic, exhibit considerable anisotropy due to reflection and refraction effects at the interfaces between the solution, the container and the surrounding air. This anisotropy represents a considerable systematic error (>25%) in some values of quantum yields reported in the literature. The degree of asymmetry in spatial distribution depends on the solution volume, the refractive index, and the degree of light scattering in the solution. Quantitative evaluation of the projected image of these sources using a video camera system indicates that a major contribution to this asymmetry is due to reflection at the meniscus. Since container frosting removes the variability due to scattering, volume changes, refractive index differences and even, to some extent, use of different types of containers, it is recommended that quantum yields and comparative measurements be determined using frosted containers and point source geometry. The container of choice is a frosted, 1 ± 7.5 cm test tube.