Fluorescence Imaging (FI) has become a major diagnostic tool in the surgical field where Indocyanine Green (ICG) is mainly used as contrast agents, as it is currently the most widely compatible, FDA-approved contrast agent. ICG stained particles emit fluorescent light when irritated by pulses of laser. For in vivo surgical imaging, however, ICG is used in either different concentrations or mixed with various viscous solutions to avoid dispersion to unnecessary areas. Depending on the mixture and the concentration, absorption levels will fluctuate and wavelengths may even shift from what is determined to be the norm. Similarly, ICG has limited capabilities in detection under certain depths. This can lead to inconsistent decisions among surgeons, further leading to reoccurring problems both during and post-surgery as detection may become faint or inaccurate. Due to its high contrast, high sensitivity and affordability, much research has been done on the properties of ICG but there currently is a lack of sufficient data on the varying shifts and absorption levels caused by different conditions. By determining and solidifying a spectrum of wavelengths that different ICG solutions emit depending on its concentration, mixture and depth, ICG detection can greatly be enhanced through better calibration. Ultimately, it will increase the effectiveness of non-invasive imaging-guided treatments and diagnostics.