We developed a custom video rate SFDI fluorescence imaging system to perform real-time fluorescence visualization with absorption minimization. The LightCrafter evaluation module (LC-EVO, Texas Instruments, Dallas, Texas) was used to generate and project patterns. Specifically, the LC-EVO worked in a stored-pattern-sequence mode, which avoided the internal gamma correction. A noncooled CCD camera (piA2400-17 gm/gc, Basler, Ahrensburg, Germany) with a fast lens (Fujinon, Valhalla, New York, HF25HA-1B) was used for image acquisition. A 490 nm LED low-pass filtered at 500 nm was used for the excitation. A 510 nm high-pass filter (BrightLine FF510, Semrock, Rochester, New York) was used for fluorescence collection. The light engine of the LC-EVO was custom modified, thus the minimal diagonal image size was reduced from 25.4 to 2.8 cm with a 2 cm working distance to achieve high intensity illumination and high spatial frequency pattern. The projection magnification and working distance can be adjusted to meet specific application requirements. In our study, the imaging area was .