Fluorescence spectroscopy has been investigated as a means of detecting oral, lung and cervical neoplasia. However, there are many issues in developing a clinical protocol for routinely making such measurements. We conducted an experiment with a simulated gynecological patient to study the effect of room light and speculum type. The speculum is needed for the medical provider to access the cervix. We investigated whether: (1) room lights present or absent had an effect and (2) the type of speculum (no speculum, coated speculum, or metal speculum) made a difference. In recent years, simulation-based medicine has gained prominence in clinical science. A simulated gynecological model was employed as a surrogate for a human cervix, which provides a physical environment similar to the clinical setting and mitigates ethical considerations. Measurements on fluorescence standards placed inside the simulated patient were made at all combinations of lighting and speculum type. Both light and the type of speculum were significant factors, and there was a statistically significant interaction between the two factors. When the room light was absent, the measurements made with either the metal speculum and the coated speculum did not exhibit significant differences, so either type of speculum may be used when the lights are absent. When the lights are present, the coated speculum shows less room light contamination in the measurements. Because the room light can contaminate the fluorescence spectroscopic measurements, making the measurement with room lights absent is highly recommended.