Cell disintegration by laser-induced transient microbubbles and its simultaneous monitoring by interferometry

Selective retina treatment (SRT) is a novel short pulsed laser therapy of several retinal diseases associated with a decreased metabolism at the retinal pigment epithelium (RPE). The range of laser pulse energies is small, in which the desired selective RPE disintegration is achieved without adverse effects to the neural retina. Thus, a real-time dosimetry control is required. We investigated a noninvasive interferometric technique able to monitor microbubble formation around the intracellular melanin granula, which is the origin of the desired RPE damage. A porcine ex vivo RPE model was irradiated by single pulses (350 ns/1.7 μs) of a neodymium: yttrium lithium fluoride laser (527 nm). The specimen was simultaneously probed by a Michelson interferometer (helium neon-laser: 633 nm) and by a hydrophone. Cell viability assays (Calcein-AM) were performed after irradiation. At threshold radiant exposure for cell death (ED₅₀ = 129±5 mJ/cm² for 350 ns; ED ₅₀=180±5 mJ/cm² for 1.7 μs), the interferometric transients changed due to microbubble formation. No major differences in the bubble dynamics were observed between both pulse durations. An algorithm to determine cell death from the interferometric transients showed less than 10% false positive or false negative results for the applied laser expositions compared to the viability assay. Interferometry is a reliable noncontact technique to monitor RPE disintegration and may serve as real-time dosimetry control during SRT.