Autofluorescence removal, multiplexing, and automated analysis methods for in-vivo fluorescence imaging.

The ability to image and quantitate fluorescently labeled markers in vivo has generally been limited by autofluorescence of the tissue. Skin, in particular, has a strong autofluorescence signal, particularly when excited in the blue or green wavelengths. Fluorescence labels with emission wavelengths in the near-infrared are more amenable to deep-tissue imaging, because both scattering and autofluorescence are reduced as wavelengths are increased, but even in these spectral regions, autofluorescence can still limit sensitivity. Multispectral imaging (MSI), however, can remove the signal degradation caused by autofluorescence while adding enhanced multiplexing capabilities. While the availability of spectral "libraries" makes multispectral analysis routine for well-characterized samples, new software tools have been developed that greatly simplify the application of MSI to novel specimens.

[1]  Michael G. Sowa,et al.  Development of visible and near-IR LCTF-based spectroscopic imaging systems for macroscopic samples , 2000, BiOS.

[2]  B. Rice,et al.  Quantitative comparison of the sensitivity of detection of fluorescent and bioluminescent reporters in animal models. , 2004, Molecular imaging.

[3]  Peter Choyke,et al.  Comparison of noninvasive fluorescent and bioluminescent small animal optical imaging. , 2003, BioTechniques.

[4]  Robert M Hoffman,et al.  Imaging tumor angiogenesis with fluorescent proteins. , 2004, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[5]  R. Weissleder,et al.  Fluorescence molecular imaging of small animal tumor models. , 2004, Current molecular medicine.

[6]  M E Dickinson,et al.  Multi-spectral imaging and linear unmixing add a whole new dimension to laser scanning fluorescence microscopy. , 2001, BioTechniques.

[7]  J. Kennedy,et al.  The nature of the chromophore responsible for naturally occurring fluorescence in mouse skin. , 1988, Journal of photochemistry and photobiology. B, Biology.

[8]  Charles L. Lawson,et al.  Solving least squares problems , 1976, Classics in applied mathematics.

[9]  Vasilis Ntziachristos,et al.  Shedding light onto live molecular targets , 2003, Nature Medicine.

[10]  A. Feldman,et al.  Noninvasive fluorescent imaging reliably estimates biomass in vivo. , 2002, BioTechniques.

[11]  K Svanberg,et al.  In vivo fluorescence imaging for tissue diagnostics. , 1997, Physics in medicine and biology.

[12]  Peter Choyke,et al.  Current Advances in Molecular Imaging: Noninvasive in Vivo Bioluminescent and Fluorescent Optical Imaging in Cancer Research , 2003, Molecular imaging.

[13]  Anton P. McCaffrey,et al.  Advancing Molecular Therapies through In Vivo Bioluminescent Imaging , 2003, Molecular imaging.

[14]  J. Kennedy,et al.  A recipe for the preparation of a rodent food that eliminates chlorophyll-based tissue fluorescence. , 1995, Journal of photochemistry and photobiology. B, Biology.

[15]  R. Pepperkok,et al.  Spectral imaging and its applications in live cell microscopy , 2003, FEBS letters.

[16]  S. Nie,et al.  In vivo cancer targeting and imaging with semiconductor quantum dots , 2004, Nature Biotechnology.

[17]  S. Libutti,et al.  Current advances in molecular imaging: noninvasive in vivo bioluminescent and fluorescent optical imaging in cancer research. , 2003 .

[18]  D L Farkas,et al.  Non-invasive image acquisition and advanced processing in optical bioimaging. , 1998, Computerized medical imaging and graphics : the official journal of the Computerized Medical Imaging Society.

[19]  D. Haaland,et al.  Multivariate Least-Squares Methods Applied to the Quantitative Spectral Analysis of Multicomponent Samples , 1985 .

[20]  Ralph Weissleder,et al.  Feasibility of in vivo multichannel optical imaging of gene expression: experimental study in mice. , 2002, Radiology.

[21]  T. Kaufman,et al.  Chemometrics-assisted simultaneous determination of atenolol and chlorthalidone in synthetic binary mixtures and pharmaceutical dosage forms , 2003, Analytical and bioanalytical chemistry.

[22]  Lars Nørgaard,et al.  Exploratory multivariate spectroscopic study on human skin , 2003, Skin research and technology : official journal of International Society for Bioengineering and the Skin (ISBS) [and] International Society for Digital Imaging of Skin (ISDIS) [and] International Society for Skin Imaging.

[23]  J. Roodenburg,et al.  Autofluorescence characteristics of healthy oral mucosa at different anatomical sites , 2003, Lasers in surgery and medicine.

[24]  Mario Winter,et al.  N-FINDR: an algorithm for fast autonomous spectral end-member determination in hyperspectral data , 1999, Optics & Photonics.

[25]  H. Shimada,et al.  Whole-body optical imaging of green fluorescent protein-expressing tumors and metastases. , 2000, Proceedings of the National Academy of Sciences of the United States of America.