The electrostatic probe-based photo-detachment technique with one or two laser beams can provide insight on the negative ion absolute densities and dynamics, respectively, in electronegative plasmas. In this work, this diagnostic is installed in the ECR-driven (2.45 GHz) negative ion source “Prometheus I” and details of the setup design along with main underlying physics are discussed. Potential origins of signal distortions and thus erroneous employment of this technique are demonstrated. Based on these observations, H- and D- negative ion densities and temperatures are measured in a reliable manner. The experiments are carried out for variable microwave power (2.45 GHz), at 1.33 Pa, unveiling two distinct kinetic equilibriums of the negative ions, in both gases, and an isotope effect.