PURPOSE.To characterize the clinical profile of femtosecond lenticule extraction (FLEx) correlated with ultrastructural analysis of the corneal interface and in vivo real-time intraocular pressure (IOP). METHODS. Prospective clinical case series with experimental studies; consecutive patients underwent FLEx at a single tertiary center over 10 months with postsurgical follow-up of 3 months. The patients were divided into three groups according to spherical equivalence (SE) (A, Ͻ Ϫ5.0 diopters [D]; B, Ն Ϫ5.00 D and Ͻ Ϫ9.00 D; and C, Ն Ϫ9.0 D). Twelve human cadaveric eyes analyzed using scanning electron microscopy after receiving FLEx; 40 rabbit eyes received FLEx with in vivo IOP measurements. The main outcome measures were refractive outcomes from study subjects; with corneal interface and IOP in experimental studies. RESULTS. Thirty-three subjects (22 females, 66.7%) underwent FLEx in both eyes (66 eyes). Mean age was 32 years (range, 21 to 46 years). Preoperative mean SE was Ϫ5.77 Ϯ 2.04 D with astigmatism of Ϫ1.03 Ϯ 0.72 D. There was a slight hyperopic shift (mean SE 0.14 Ϯ 0.53 D); 94% achieved uncorrected visual acuity Ն20/25 3 months postoperatively. Refractive stability was achieved within 1 month (P Ͻ 0.001). Ultrastructurally, the smoothness of the corneal interface was independent of ablation depth (mean irregularity scores A, B, C: 8.8 Ϯ 0.6, 10.3 Ϯ 0.4, 8.7 Ϯ 0.6, respectively; P ϭ 0.88). The increase in IOP during FLEx was similar to that in femtosecond (FS)-LASIK, albeit a twofold duration of raised IOP in FLEx (P Ͻ 0.001). CONCLUSIONS. These results suggest that FLEx is predictable and effective in treating myopia and myopic astigmatism. Experimental studies support the early clinical results and safety of this procedure. (Invest Ophthalmol Vis Sci. 2012;53: 1414 -1421) DOI:10.1167/iovs.11-8808 T he femtosecond (FS) laser is a near-infrared neodymiumdoped yttrium aluminum garnet (ND:YAG) laser that photodisrupts the cornea with surgical precision through plasma cleavage of stromal lamellae.1 The first commercial ophthalmic FS laser was introduced into the market in 2001 for laser in situ keratomileusis (LASIK) flaps. Since then, significant improvements in laser energy profiles, spatial resolution, and faster laser speeds have occurred.2 FS lasers have thus made a significant impact on refractive surgery by enabling nonmechanical creation of a corneal flap during LASIK. The FS laser offers several advantages over manual microkeratomes including increased precision, reduced incidence of flap complications, and the ability to cut thinner flaps without the risk of buttonhole formation.3,4 Moreover, newer generation FS lasers have reduced problems such as transient lightsensitivity syndrome and interference by cavitation bubbles. 5,6