Cherenkov radiation has emerged as a novel source of light with a number of applications in the biomedical sciences. It’s unique properties, including its broadband emission spectrum, spectral weighting in the ultraviolet and blue wavebands, and local generation of light within a given tissue have made it an attractive source of light for techniques ranging from widefield imaging to oximetry and phototherapy. To help guide the future development of this field in the context of molecular imaging, quantitative estimates of the light fluence rates of Cherenkov radiation from a number of radionuclide and external radiotherapy beams in tissue was explored for the first time. Using Monte Carlo simulations, these values were found to be on the order of 0.1 – 1 nW/cm2 per MBq/g for radionuclides and 1 – 10 μW/cm2 per Gy/sec for external radiotherapy beams, dependent on the given waveband and optical properties. For phototherapy applications, the total light fluence was found to be on the order of nJ/cm2 for radionuclides, and mJ/cm2 for radiotherapy beams. To validate these findings, experimental validation was completed with an MV x-ray photon beam incident onto a tissue phantom, confirming the magnitudes of the simulation values. The results indicate that diagnostic potential is reasonable for Cherenkov excitation of molecular probes, but phototherapy may remain elusive at these relatively low fluence values.