Various neurologic disorders such as chronic pain or Alzheimer's disease are known to be associated with abnormal imbalances in chloride homeostasis, hence the importance of developing better chloride indicators. SuperClomeleon is a fluorescent chloride indicator based on Förster Resonance Energy Transfer (FRET). In an effort to improve SuperClomeleon's efficiency, genetic mutations were made to modify the linker between the donor fluorophore (Cerulean) and its acceptor (eYFP) to increase the dynamic range of chloride response. The hypothesis suggesting that a modified linker, such as that presented in an article by Murakoshi et al. (Nature Scientific Reports, 2017), could potentially improve FRET efficiency is investigated. FRET efficiency is measured and calculated using two methods, both imaged through two-photon microscopy; either via a ratio between the intensity of emitted light in the spectral area of the acceptor fluorescence over that of the donor fluorescence, or via Fluorescence Lifetime Imaging Microscopy (FLIM) to measure the lifetime of the donor fluorophore. In both cases, microscopy is performed on rat cortical neuronal primary cultures. Using intracellular chloride concentration calibration protocols, the dynamic range of the variants are explored using a combination of multiexponential fits and phasor plot analysis techniques. This characterization of a modified genetic chloride indicator aims to provide improved fluorescent tools to monitor chloride in vivo and allow complex assessment of ion homeostasis in brain disorders.