A 110 × 64 pixel CMOS image sensor integrated with pixelated spectral interference filters is used for image guided surgery. The sensor operates with short exposure times for capturing visible light using blue, green, and red pixelated interference filters, and long exposure time for capturing near-infrared fluorescence with a co-located, pixelated NIR filter. The sensor was used to successfully detect the fluorescence of ICG accumulation in sentinel lymph node on patients with breast cancer.