Background Subthreshold microsecond pulsing laser is an increasingly common treatment approach for central serous chorioretinopathy. However, there is no literature available on the safety of microsecond laser using different fluence settings in this disease. While many publications can be obtained from conventional microsecond pulsing lasers, few parameter sets are published with the navigated microsecond pulsing laser. Therefore, this study aims to investigate the safety of different parameter sets in subthreshold microsecond pulsing laser treatments. Methods In this retrospective chart review, consecutive patients with central serous chorioretinopathy (> 3 months duration of symptoms) treated with navigated subthreshold microsecond pulsing laser and a follow up of at least five months after microsecond laser application were included. For each patient, the treatment parameters, plan layout, and adverse events related to laser were evaluated. Secondary outcomes included best-corrected visual acuity and anatomical improvements (central retinal thickness). Results One hundred and one eyes were included in the observation and followed for a mean of 10 months (range 5–36). Although a larger range of parameter sets and fluence settings have been used, no patient demonstrated adverse events from navigated microsecond pulsing laser. While 88% of the cases demonstrated stability, 13 cases lost five or more letters due to the persistence of the subretinal fluid. In mean, a best-corrected visual acuity improvement of 0.07logMar (± 0.2) was seen (p = 0.02). In 51% of the patients, a statistically significant improvement of the central retinal thickness was noted at the last follow-up with a mean thickness reduction of 70 µm (± 143) (p < 0.01). Conclusion In conclusion, none of the used parameter sets lead to tissue damage (when using a cautious titration) and, in summary, lead to an improvement in subretinal fluid and improvement in visual acuity. However, further prospective studies are needed to correctly identify the dependency of the treatment strategy on the outcome criteria.