Application Overview

Optical imaging is a fast, sensitive, and cost effective way to image and track molecules in small animals. There are two main types of optical imaging:

• Bioluminescent imaging requires genetic alteration of cells with a reporter gene (e.g. luciferase). After injection of a substrate such as luciferin, substrate oxidation occurs and emitted photons can be detected by a camera.

• Fluorescent imaging is able to use native, unaltered cells for the visualization of molecular events in the animal. A fluorescently labeled targeting agent (peptide, protein, cell, etc.) is injected into the animal, where it will either be cleared from the animal’s circulation over time or retained by binding to a specific target. Upon excitation with a light source, the fluorescent dye will emit photons that are collected by a sensitive detector.

 

Fluorescent optical probes (also called targeting agents) are diverse, and may include peptides, proteins, antibodies, or small molecules that are covalently labeled with fluorescent dyes.

The best choice of optical probe depends on your research goals. You may wish to:

• Target a cell surface protein, such as a receptor or transporter

• Illuminate a structural feature, such as bone

• Visualize blood flow and pooling in the vasculature, for biodistribution or angiogenesis studies

• Track the biodistribution of labeled probes, cells, viruses, etc.

• Develop and label your own probes

 

Near-infrared dyes, such as IRDye^® fluorophores, and carefully optimized hardware are critical for high-performance optical imaging.

• Near-infrared fluorophores exploit the spectral region where light absorption and scatter properties of tissue are most advantageous¹. This enhances penetration depth (access of excitation light to the fluorophore) and escape of emitted fluorescence from the animal to reach the detector.

• Laser illumination delivers very intense excitation light of the correct wavelength, generating the brightest possible signal from the fluorescent agent.

• Intrinsic autofluorescence from animal tissue can mask the signal from optical probes. In the NIR spectral region, autofluorescence is dramatically lowered ^2,3.

¹ Tsien, R. Science. Vol. 324, May 8, 2009

² Hawrysz, DJ and Sevick-Muraca, EM. Neoplasia 2(5):388–417 (2000)

³ Frangioni, JV. Curr Opin Chem Biol. 7(5):626-34 (2003)

⁴ Adams, KE, et al. J Biomed Opt. 12(2):024017 (2007)

Webinars

• Why Image in the NIR?

  • “Advances in In Vivo Imaging: Near-Infrared Optical Imaging of Mice”
    Jeff Harford, LI-COR

  • “Instrumentation and Imaging Considerations”
    Eva Sevick-Muraca, Baylor College of Medicine

  • “In vivo imaging of prostate cancer using an IRDye^® 800CW EGF Optical Probe”
    Melanie Simpson, University of Nebraska – Lincoln

  • “Comparison of visible and near-infrared wavelength-excitable fluorescent dyes for molecular imaging of cancer”
    Eva Sevick-Muraca and Shi Ke, Baylor College of Medicine

• Development of imaging agents

  • “Systematic Evaluation and Use of Targeted IRDye labeled Optical Contrast Agents”
    Mike Olive, LI-COR

  • “Systematic Evaluation of Targeted IRDye Labeled Optical Imaging Agents”
    Joy Kovar, LI-COR

  • “Near Infrared Fluorescent Approaches to Cell-Based Assays and Small Animal Imaging”
    Amy Geschwender, LI-COR

• Translational Imaging

  • “Nodal Staging of HER2 Positive Breast Cancer with Dual-Labeled Trastuzuman Based Imaging Agent”
    Lakshmi Sampath, Baylor College of Medicine

• Targeted Imaging

  • "Illuminating Prostate Cancer Progression"
    Melanie Simpson, University of Nebraska - Eppley Cancer Center

  • “IRDye^® 800 2DG: A Broadly Applicable Tumor Imaging Agent for Preclinical Research”
    Mike Olive, LI-COR

• Five-Part Educational Webinar Series: Optical Imaging

  • “Part 1: A Technology Introduction to Optical Imaging”
    Jeff Harford, Senior Product Manager, LI-COR

  • “Part 2: A Technical Overview of in vivo Optical Imaging Agents & Targets”
    Peter Johnson, Senior Field Applications Scientist, LI-COR

  • “Part 3: A Technical Overview of Optical Imaging: Light Sources & Target Illumination”
    Rex Peterson, Engineering Product Support Manager, LI-COR

  • “Part 4: A Technical Overview of Optical Imaging: Dye Absorption & Fluorescence”
    Rex Peterson, Engineering Product Support Manager, LI-COR

  • “Part 5: A Technical Overview of Optical Imaging: Light Collection, Imaging Display & Dynamic Range”
    Rex Peterson, Engineering Product Support Manager, LI-COR

PubMed

Publishing with LI-COR data?

• Why image in the near-infrared?

  • Adams, KE et al.
    Comparison of visible and near-infrared wavelength-excitable fluorescent dyes for molecular imaging of cancer.
    J Biomed Opt. 12(2):024017 (2007)

  • Kovar, J et al.
    A systematic approach to the development of fluorescent contrast agents for optical imaging of mouse cancer models.
    Anal Biochem. 367():1-12 (2007)

  • Osterman, H and Schutz-Geschwender, A.
    Seeing beyond the visible with IRDye infrared dyes
    LI-COR Biosciences (2007)

  • Olive, DM.
    Near infrared technology and optical agents for molecular imaging
    LI-COR Biosciences(2006)

• Development of imaging agents

  • Kovar, J et al.
    Characterization and performance of a near-infrared 2-deoxyglucose optical imaging agent for mouse cancer models.
    Anal Biochem.384(2): 254-62 (2009)

  • Kovar, J et al.
    A systematic approach to the development of fluorescent contrast agents for optical imaging of mouse cancer models.
    Anal Biochem. 367(1):1-12 (2007)

  • Kovar, J et al.
    Purification method directly influences effectiveness of an epidermal growth factor-coupled targeting agent for noninvasive tumor detection in mice.
    Anal Biochem. 361(1):47-54 (2007)

  • Kovar, J. et al.
    EGF-IRDye 800CW: in vitro and in vivo characterization as a biomarker for optical fluorescent imaging of tumor growth kinetics.
    Poster presentation, SMI Annual Meeting (2005)

• Targeted Imaging

  • Banerjee, S et al.
    Antitumor activity of gemcitabine and oxaliplatin is augmented by thymoquinone in pancreatic cancer
    Cancer Res. 69(13):5575-83 (2009)

  • Banerjee, S et al.
    3,3'-Diindolylmethane enhances chemosensitivity of multiple chemotherapeutic agents in pancreatic cancer
    Cancer Res. 69(13):5592-600 (2009)

  • Mondal, S et al.
    Functional Blocking Monoclonal Antibodies against IL-12p40 Homodimer Inhibit Adoptive Transfer of Experimental Allergic Encephalomyelitis.
    J Immunol. 182(8):5013-23 (2009)

  • Rosenzweig, HL et al.
    Activation of nucleotide oligomerization domain 2 exacerbates a murine model of proteoglycan-induced arthritis.
    J Leukoc Biol. 85(4):711-8 (2009)

  • Roy, EJ et al.
    Imaging membrane intercalating near infrared dyes to track multiple cell populations.
    J Immunol Methods. 348(1-2):18-29 (2009)

  • Bell, LN et al.
    A central role for hepatocyte growth factor in adipose tissue angiogenesis.
    Am J Physiol Endocrinol Metab. 294(2):E336-44 (2008)

  • Duysen, EG and Lockridge, O.
    Whole body and tissue imaging of the butyrylcholinesterase knockout mouse injected with near infrared dye labeled butyrylcholinesterase.
    Chem Biol Interact. 175(1-3):119-24 (2008)

  • He, X et al.
    Cytotoxicity of Paclitaxel in Biodegradable Self-Assembled Core-Shell Poly (Lactide-Co-Glycolide Ethylene Oxide Fumarate) Nanoparticles.
    Pharm Res. 25(7):1552-62 (2008)

  • Senf, SM et al.
    Hsp70 overexpression inhibits NF-kappaB and Foxo3a transcriptional activities and prevents skeletal muscle atrophy.
    FASEB J. 22(11):3836-45 (2008)

  • Wang, GJ et al.
    Thymus Exosomes-Like Particles Induce Regulatory T Cells.
    J Immunol. 181(8):5242-8 (2008)

  • Kovar, J et al.
    GLUT Family Transporters Involvement in Cellular Uptake of IRDye 800CW 2-Deoxyglucose.
    Poster presentation, SMI Annual Meeting (2008)

  • Kovar, J et al.
    Purification method directly influences effectiveness of an epidermal growth factor-coupled targeting agent for noninvasive tumor detection in mice.
    Anal Biochem. 361(1):47-54 (2007)

  • Sampath, L et al.
    Dual-labeled trastuzumab-based imaging agent for the detection of human epidermal growth factor receptor 2 overexpression in breast cancer.
    J Nucl Med. 48(9):1501-10 (2007)

  • Kovar, J et al.
    Effective bone labeling for in vivo NIR noninvasive imaging in nude mice.
    Poster presentation, Joint Molecular Imaging Conference (2007)

  • Roy, A et al.
    Myelin basic protein-primed T cells induce neurotrophins in glial cells via alphavbeta3 integrin.
    J Biol Chem. 282(44):32222-32 (2007)

  • Kovar, J et al.
    Hyaluronidase expression induces prostate tumor metastasis in an orthotopic mouse model.
    Am J Pathol. 169(4):1415-26 (2006)

  • Kovar, J et al.
    Monitoring progression of prostate tumors in mice by receptor-targeted near infrared optical imaging.
    Poster presentation, In Vivo Molecular Imaging Annual Meeting (2005)

• Biodistribution

  • see Biodistribution

  • Roy, EJ et al.
    Imaging membrane intercalating near infrared dyes to track multiple cell populations.
    J Immunol Methods. 348(1-2):18-29 (2009)

  • Kovar, J et al.
    Characterization and performance of a near-infrared 2-deoxyglucose optical imaging agent for mouse cancer models.
    Anal Biochem.384(2): 254-62 (2009)

  • Duysen, EG and Lockridge, O.
    Whole body and tissue imaging of the butyrylcholinesterase knockout mouse injected with near infrared dye labeled butyrylcholinesterase.
    Chem Biol Interact. 175(1-3):119-24 (2008)

  • Kovar, J et al.
    GLUT Family Transporters Involvement in Cellular Uptake of IRDye 800CW 2-Deoxyglucose.
    Poster presentation, SMI Annual Meeting (2008)

  • Kovar, J et al.
    Effective bone labeling for in vivo NIR noninvasive imaging in nude mice.
    Poster presentation, Joint Molecular Imaging Conference (2007)

  • Kovar, J et al.
    Purification method directly influences effectiveness of an epidermal growth factor-coupled targeting agent for noninvasive tumor detection in mice.
    Anal Biochem. 361(1):47-54 (2007)

  • Kovar, J et al.
    Hyaluronidase expression induces prostate tumor metastasis in an orthotopic mouse model.
    Am J Pathol. 169(4):1415-26 (2006)

  • Kovar, J. et al.
    EGF-IRDye 800CW: in vitro and in vivo characterization as a biomarker for optical fluorescent imaging of tumor growth kinetics.
    Poster presentation, SMI Annual Meeting (2005)

  • Kovar, J et al.
    Monitoring progression of prostate tumors in mice by receptor-targeted near infrared optical imaging.
    Poster presentation, In Vivo Molecular Imaging Annual Meeting (2005)

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