Planar fluorescence imaging using normalized data.
Fluorescence imaging of tissues has gained significant attention in recent years due to the emergence of appropriate reporter technologies that enable noninvasive sensing of molecular function in vivo. Two major approaches have been used so far for fluorescence molecular imaging, i.e., epi-illumination (reflectance) imaging and fluorescence molecular tomography. Transillumination is an alternative approach that has been employed for imaging tissues in the past and could be similarly beneficial for fluorescence molecular imaging. We investigate data normalization schemes in reflectance and transillumination mode and experimentally demonstrate that normalized transillumination offers significant advantages over planar reflectance imaging and over nonnormalized methods. Our observations, based on phantoms and on postmortem and in vivo mouse measurements display image quality improvement, superior depth sensitivity, and improved imaging accuracy over the nonnormalized methods examined. Normalized planar imaging retains implementation simplicity and could be used to improve on standard fluorescence reflectance imaging and as a simplified alternative to the more integrated and accurate tomographic methods.
Ntziachristos
Vasilis
V
Massachusetts General Hospital & Harvard Medical School, Center for Molecular Imaging Research, Laboratory for Biooptics and Molecular Imaging, Charlestown, Massachusetts 02129, USA. vasilis@helix.mgh.harvard.edu
Turner
Gordon
G
Dunham
Joshua
J
Windsor
Stephen
S
Soubret
Antoine
A
Ripoll
Jorge
J
Shih
Helen A
HA
D000465:Algorithms; D000818:Animals; D003627:Data Interpretation, Statistical; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008325:Mammary Neoplasms, Experimental / Q000473:pathology*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D019047:Phantoms, Imaging; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D051598:Whole Body Imaging / Q000379:methods*