Date of Degree

8-2025

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Program

Vision Science

Advisor

Brian K. Foutch

Advisor

Jeffrey C. Rabin

Advisor

Michelle T. Aaron

Advisor

Michael L. Denton

Abstract

Excess optical radiation can injure the retina through either rapid heat deposition (photothermal damage) or cumulative photo‑oxidative stress (photochemical damage), yet current laser‑safety limits treat these mechanisms as independent. To test that assumption, this dissertation integrated systematic wavelength, duration and temperature‑controlled laser exposures with live/dead fluorescence imaging in a melanin‑loaded hTERT‑RPE1 monolayer model. First, concurrent 447 nm (blue) and 2 µm (infra‑red) beams delivered for 200 s demonstrated a strong thermal–photochemical synergy: each beam alone was sub‑threshold (≤25 % lethality) but together produced 100 % RPE death without exceeding 50 °C, proving that modest heating markedly lowers the photon dose required for photochemical injury. Second, continuous‑wave thresholds were mapped at 413, 447 and 488 nm over 0.25–400 s. All three wavelengths exhibited two‑segment ED₅₀ curves: steep, time‑dependent irradiance at short exposures (photothermal damage) followed by wavelength‑specific plateaus that obey intensity–time reciprocity (photochemical damage). Plateau radiant‑exposure values increased with wavelength (0.94, 2.1 and 4.2 kJ cm⁻², respectively), and the photothermal–photochemical crossover shifted later in time from 413 nm (~60–100 s) to 488 nm (>120 s), confirming a weighted action spectrum with shorter wavelengths. Together these results show that (i) sub‑threshold heating can amplify blue‑light photochemical toxicity, and (ii) photochemical reciprocity limits are wavelength‑dependent rather than fixed. Although derived from an isolated RPE layer in vitro, the data indicate that ANSI Z136.1 dual‑limit exposure criteria may underestimate retinal risk when moderate temperature rises coincide with short‑wavelength light. The work provides quantitative benchmarks for refining mixed‑mechanism safety models and guides future in vivo validation of wavelength‑ and temperature‑dependent retinal hazards.

Journal of Biomedical Optics Permission 20250627.pdf (235 kB)
Permission to use data from another study for Chapter 3

thesis-dissertation page template VEditedbyUIW V2 comments.pdf (2956 kB)
Edits made by editor with my comments that addresses each edit

Available for download on Tuesday, June 30, 2026

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