Spectral near-infrared fluorescence imaging of curved surfaces using projection reconstruction algorithms

In vivo spectral fluorescence imaging has made it possible to non-invasively visualize superficial curved structures as well as structures deep to the skin. However, the defocus created by blurring has been an obstacle to creating anatomically interpretable surface images. Herein we present a methodology to correct for blurring induced by curved structures during spectral fluorescence imaging using signal intensity projection algorithms. In a phantom and an animal model in which the lymphatic system was visualized after the interstitial injection of quantum dots with emission spectra in the near-infrared (NIR) range, the planes of focus were sequentially adjusted to obtain a z-stack of images which contains images acquired from multiple focal points. Maximum, minimum, median and average intensity projections were applied to the resulting images. Using the phantom, the minimum and the median intensity projection images demonstrated improved deblurring whereas during in vivo imaging the median intensity projection images more clearly visualized important structures than did the other projection techniques. Image stacking with subsequent application of appropriate projection techniques provides a simple method for deblurring in vivo optical images obtained from curved surfaces, thus improving their anatomic resolution.