Abstract

We present a fast scanning fluorescence optical tomography system for imaging the kinetics of probe distributions through out the whole body of small animals. Configured in a plane parallel geometry, the system scans a source laser using a galvanometer mirror pair (τswitch~1ms) over flexible source patterns, and detects excitation and emission light using a high frame rate low noise, 5 MHz electron multiplied charge-coupled device (EMCCD) camera. Phantom studies were used to evaluate resolution, linearity, and sensitivity. Time dependent (δt=2.2 min.) in vivo imaging of mice was performed following injections of a fluorescing probe (indocyanine green). The capability to detect differences in probe delivery route was demonstrated by comparing an intravenous injection, versus an injection into a fat pocket (retro orbital injection). Feasibility of imaging the distribution of tumor-targeted molecular probes was demonstrated by imaging a breast tumor-specific near infrared polypeptide in MDA MB 361 tumor bearing nude mice. A tomography scan, at 24 hour post injection, revealed preferential uptake in the tumor relative to surrounding tissue.

© 2005 Optical Society of America

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Acad. Radiol.

Ntziachristos, V., C. Bremer, C. Tung and R. Weissleder, "Imaging cathepsin B up-regulation in HT-1080 tumor models using fluorescence-mediated molecular tomography (FMT)," Acad. Radiol. 9, 323-325 (2002).
[CrossRef]

Appl. Opt.

Cancer Res.

Eccles, S.A., W.J. Court, G.A. Box, C.J. Dean and R.G. Melton, "Regression of Established Breast-Carcinoma Xenografts with Antibody-Directed Enzyme Prodrug Therapy against C-Erbb2 P185," Cancer Res. 54, 5171-5177 (1994).
[PubMed]

Current Molecular Medicine

Graves, E.E., R. Weissleder and V. Ntziachristos, "Fluorescence molecular imaging of small animal tumor models," Current Molecular Medicine 4, 419-430 (2004).
[CrossRef] [PubMed]

IEEE Compu. Sc. & Engg.

Barbour, R.L., H.L. Graber, J.W. Chang, S.L.S. Barbour, P.C. Koo and R. Aronson, "MRI-guided optical tomography: Prospects and computation for a new imaging method," IEEE Compu. Sc. & Engg. 2, 63-77 (1995).
[CrossRef]

IEEE Trans. Med. Imaging

Schulz, R.B., J. Ripoll and V. Ntziachristos, "Experimental fluorescence tomography of tissues with noncontact measurements," IEEE Trans. Med. Imaging 23, 492-500 (2004).
[CrossRef] [PubMed]

Invest. Radiol.

Achilefu, S., R.B. Dorshow, J.E. Bugaj and R. Rajagopalan, "Novel receptor-targeted fluorescent contrast agents for in vivo tumor imaging," Invest. Radiol. 35, 479-485 (2000).
[CrossRef] [PubMed]

J. Biomed. Opt.

Bugaj, J.E., S. Achilefu, R.B. Dorshow and R. Rajagopalan, "Novel fluorescent contrast agents for optical imaging of in vivo tumors based on a receptor-targeted dye-peptide conjugate platform," J. Biomed. Opt. 6, 122-133 (2001).
[CrossRef] [PubMed]

Srinivasan, S., B.W. Pogue, H. Dehghani, S.D. Jiang, X.M. Song and K.D. Paulsen, "Improved quantification of small objects in near-infrared diffuse optical tomography," J. Biomed. Opt. 9, 1161-1171 (2004).
[CrossRef] [PubMed]

J. Opt. Soc. Am. A

Med. Phys.

Intes, X., J. Ripoll, Y. Chen, S. Nioka, A.G. Yodh and B. Chance, "In vivo continuous-wave optical breast imaging enhanced with Indocyanine Green," Med. Phys. 30, 1039-1047 (2003).
[CrossRef] [PubMed]

Culver, J.P., R. Choe, M.J. Holboke, L. Zubkov, T. Durduran, A. Slemp, V. Ntziachristos, B. Chance and A.G. Yodh, "Three-dimensional diffuse optical tomography in the parallel plane transmission geometry: evaluation of a hybrid frequency domain/continuous wave clinical system for breast imaging," Med. Phys. 30, 235-247 (2003).
[CrossRef] [PubMed]

Godavarty, A., C. Zhang, M.J. Eppstein and E.M. Sevick-Muraca, "Fluorescence-enhanced optical imaging of large phantoms using single and simultaneous dual point illumination geometries," Med. Phys. 31, 183-190 (2004).
[CrossRef] [PubMed]

Graves, E.E., J. Ripoll, R. Weissleder and V. Ntziachristos, "A submillimeter resolution fluorescence molecular imaging system for small animal imaging," Med. Phys. 30, 901-911 (2003).
[CrossRef] [PubMed]

Ntziachristos, V. and R. Weissleder, "Charge-coupled-device based scanner for tomography of fluorescent near-infrared probes in turbid media," Med. Phys. 29, 803-809 (2000).
[CrossRef]

Pogue, B.W., C. Willscher, T.O. McBride, U.L. Osterberg and K.D. Paulsen, "Contrast-detail analysis for detection and characterization with near-infrared diffuse tomography," Med. Phys. 27, 2693-2700 (2000).
[CrossRef]

Nat. Med.

Ntziachristos, V., C.H. Tung, C. Bremer and R. Weissleder, "Fluorescence molecular tomography resolves protease activity in vivo," Nat. Med. 8, 757-760 (2002).
[CrossRef] [PubMed]

Opt. Exp.

Durduran, T., J.P. Culver, M.J. Holboke, X.D. Li, L. Zubkov, B. Chance, D.N. Pattanayak and A.G. Yodh, "Algorithms for 3D localization and imaging using near-field diffraction tomography with diffuse light," Opt. Exp. 4, 247-262 (1999).
[CrossRef]

Opt. Lett.

Optics Letters

Li, X.D., T. Durduran, A.G. Yodh, B. Chance and D.N. Pattanayak, "Diffraction tomography for biochemical imaging with diffuse- photon density waves," Optics Letters 22, 573-575 (1997).
[CrossRef] [PubMed]

Photochem. Photobiol.

Gurfinkel, M., A.B. Thompson, W. Ralston, T.L. Troy, A.L. Moore, T.A. Moore, J.D. Gust, D. Tatman, J.S. Reynolds, B. Muggenburg, K. Nikula, R. Pandey, R.H. Mayer, D.J. Hawrysz and E.M. Sevick-Muraca, "Pharmacokinetics of ICG and HPPH-car for the detection of normal and tumor tissue using fluorescence, near-infrared reflectance imaging: A case study," Photochem. Photobiol. 72, 94-102 (2000).
[CrossRef] [PubMed]

Phys. Med. Biol.

Pogue, B.W., M.S. Patterson, H. Jiang and K.D. Paulsen, "Initial Assessment of a Simple System For Frequency-Domain Diffuse Optical Tomography," Phys. Med. Biol. 40, 1709-1729 (1995).
[CrossRef] [PubMed]

Godavarty, A., M.J. Eppstein, C.Y. Zhang, S. Theru, A.B. Thompson, M. Gurfinkel and E.M. Sevick-Muraca, "Fluorescence-enhanced optical imaging in large tissue volumes using a gain-modulated ICCD camera," Phys. Med. Biol. 48, 1701-1720 (2003).
[CrossRef] [PubMed]

Phys. Rev. E

Gonatas, C.P., M. Ishii, J.S. Leigh and J.C. Schotland, "Optical Diffusion Imaging Using a Direct Inversion Method," Phys. Rev. E 52, 4361-4365 (1995).
[CrossRef]

Proc. National Academy of Sciences

Ntziachristos, V., E.A. Schellenberger, J. Ripoll, D. Yessayan, E. Graves, A. Bogdanov, L. Josephson and R. Weissleder, "Visualization of antitumor treatment by means of fluorescence molecular tomography with an annexin V-Cy5.5 conjugate," Proc. National Academy of Sciences of the United States of America 101, 12294-12299 (2004).
[CrossRef]

Ntziachristos, V., A.G. Yodh, M. Schnall and B. Chance, "Concurrent MRI and diffuse optical tomography of breast after indocyanine green enhancement," Proc. National Academy of Sciences of the United States of America 97, 2767-2772 (2000).
[CrossRef]

Radiol.

Bremer, C., S. Bredow, U. Mahmood, R. Weissleder and C.H. Tung, "Optical imaging of matrix metalloproteinase-2 activity in tumors: Feasibility study in a mouse model," Radiol. 221, 523-529 (2001).
[CrossRef]

Tech. in Cancer Research & Treatment

Pogue, B.W., S.L. Gibbs, B. Chen and M. Savellano, "Fluorescence imaging in vivo: Raster scanned point-source imaging provides more accurate quantification than broad beam geometries," Tech. in Cancer Research & Treatment 3, 15-21 (2004).

Topics in Current Chemistry

Achilefu, S. and R.B. Dorshow, "Dynamic and Continuous Monitoring of Renal and Hepatic Functions with Exogenous Markers," Topics in Current Chemistry. Springer-Verlag: Berlin Heidelberg (2002).

Other

Kak, A.C. and M. Slaney, "Principles of Computerized Tomographic Imaging," New York: IEEE Press (1988).

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Figures (8)

Fig. 1.
Fig. 1.

Fast scanning fluorescence tomography system. The mouse subject is suspended and held in light compression between two movable windows (W1 and W2). Light from a laser diode at 785 nm (LD) is collimated and passes through a 95/5 beam splitter (BS). A reference Photodiode (PD) collects 5% of the beam. The main 95% beam passes through lens (L1) into a XY galvo scanning system (XYGal). The mirror pair scans the beam onto the illumination window (W1) of the imaging tank. Light emitted from W2 is detected by an EMCCD via a filter (F1) and lens system (L2).

Fig. 2.
Fig. 2.

Small animal fluorescence DOT system. The mouse subject is suspended between two movable windows. Light from a near infrared laser diode (785 nm) is scanned by a mirror pair on to the illumination window of the imaging tank. Fluorescing light emitted from the opposing window is detected by a lens coupled EMCCD Camera after passing through a filter that rejects the excitation light. CCD images are acquired for each source position. The full data set is then inverted to provide tomographic images of the concentration of exogenous fluorescing agents.

Fig. 3.
Fig. 3.

Calibration and Sensitivity. Known concentration -vs- raw reconstruction values for titrations of ICG in 3 mm tube phantoms. The data establish a calibration coefficient, and establish the region of linear response.

Fig. 4.
Fig. 4.

Resolution vs depth. a) Phantom measurement set-up. Depth of the phantom ztarget is measured from the detection plane. b) A xz plane slice at y=0 from a reconstructed volume of two 1.6mm diameter tubes (with 0.1 µM ICG). c) Full width half maximum (FWHM) Vs. Depth measured from the detection plane.

Fig. 5.
Fig. 5.

Biodistribution of indocynaine green (ICG) 3 min after retro orbital injection. 2D slices obtained from the 3D reconstruction are shown at various depths, ztarget, from the detection plane. The arrows indicate the site of ICG administration (Retro Orbital) ztarget=2.5, kidneys for ztarget=4.5, and the liver ztarget=10.5. Note the localization of ICG in the liver as compared to the kidneys 3 minutes after the probe delivery.

Fig. 6.
Fig. 6.

Volumes of interest used for head, shoulder, liver, and kidneys in the x-y plane.

Fig. 7.
Fig. 7.

Temporal kinetics of ICG. a) Time course of ICG distribution for retro-orbital injection in ROI’s for liver, kidney, head and shoulder. b) Time course comparison for ICG between IV and Retro Orbital delivery.

Fig. 8.
Fig. 8.

Representative slices from a 3D tomographic reconstruction of a nude mouse with a subcutaneous breast-specific human breast cancer xenograft MDA MD 361. a) a xy slice parallel to the detector plane at a depth of z=2.5mm and b) a xz slice extending from the source plane to detector plane at y=12mm.

Equations (3)

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y i = [ Φ ( r s ( i ) , r d ( i ) , λ emi ) θ f Φ o ( r s ( i ) , r d ( i ) , λ exc ) Φ o ( r s ( i ) , r d ( i ) , λ exc ) ]
A i , j = S o v h 3 D o G ( r s ( i ) , r j , λ exc ) G ( r j , r d ( i ) , λ emi ) G ( r s ( i ) , r d ( i ) , λ exc )
x j = N j

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