Disclosed herein are imaging systems and methods for simultaneous near-infrared light and visible light imaging of a sample comprising: a detector to form a fluorescence image of the sample and a visible image of the sample； a light source configured to emit infrared light to induce fluorescence from the sample； and a plurality of optics arranged to direct the infrared light toward the sample and form the fluorescence image of the sample and the visible light image of the sample on the detector, wherein the infrared light is directed to the sample substantially coaxially with fluorescence light received from the sample in order to decrease shadows.本文揭露了用於同時對樣本進行近紅外光和可見光成像之成像系統及方法，包括：檢測器，該檢測器用於形成該樣本的螢光圖像以及該樣本的可見圖像；光源，該光源被配置成用於發射紅外光以便從該樣本誘發螢光；以及多個光學器件，該多個光學器件被安排成用於將該紅外光朝向該樣本引導並且在該檢測器上形成該樣本的螢光圖像以及該樣本的可見光圖像，其中該紅外光與從該樣本接收的螢光基本上同軸地引導至該樣本以便減少陰影。
