Pump-probe optical coherence tomography using indocyanine green as a contrast agent

Use of indocyanine green (ICG), an FDA-approved dye, in a pump-probe scheme for optical coherence tomography (OCT) is reported. Aqueous solutions of ICG are not stable, i.e., the dye degrades over time especially in the presence of light. Addition of protein such as bovine serum albumin (BSA) stabilizes the ICG; however, when exposed to high intensity illumination, the dye still degrades. Moreover, the photodegradation is permanent and occurs swiftly if the illumination band corresponds to the ICG absorption peak. The permanence of the photobleached state illustrates that ICG photobleaching phenomenon has great potential to achieve contrast in OCT. ICG solutions with 50 micromolar concentration were prepared in water, 1% BSA, and 0.8% agarose to study the dynamics of the dye for different illumination intensity levels. In addition, different molar concentrations of ICG in water were studied for fixed illumination intensity. In each case, probability of photobleaching, defined as the ratio of the total photobleached ICG molecules to the total photons absorbed by the ground-state molecules, is evaluated to characterize the photobleaching phenomenon in ICG. We also demonstrate ICG-based pump-probe MCOCT imaging by mapping the distribution of ICG in a stage 54 Xenopus laevis.