Our goal was to assess whether quantitative fetal fibronectin (qfFN) is associated with spontaneous preterm birth (sPTB) after laser surgery for twin-twin transfusion syndrome (TTTS). qfFN was collected within 24 hours before and after laser surgery. Aims were: (1) To determine if qfFN changed with operative fetoscopy; and (2) To estimate the number of patients needed to study the predictive value of qfFN for sPTB <28 and <32 weeks. Results are reported as median (range). Among 49 patients, there was no net difference in qfFN levels after laser surgery [0.0 ng/mL (-37 to +400), p = 0.6041]. However, patients with a qfFN increase >10 ng/mL were 19 times more likely to undergo sPTB at <28 weeks (OR = 19.5). We determined that 383 and 160 patients would be needed to achieve adequate statistical power for qfFN to be predictive of sPTB at a GA <28 weeks and <32 weeks, respectively. In conclusion, laser surgery did not alter the qfFN level within the entire cohort, but qfFN may be useful in identifying a subset of patients at increased risk of preterm delivery.