In-vivo confocal real-time mini-microscopy in animal models of human inflammatory and neoplastic diseases

 

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Background and study aims: Although various improvements in tissue imaging modalities have recently been achieved, in-vivo molecular and subsurface imaging in the field of gastroenterology remains a technical challenge. In this study we evaluated a newly developed, handheld, miniaturized confocal laser microscopy probe for real-time in-vivo molecular and subsurface imaging in rodent models of human disease.

Materials and methods: The minimicroscope uses a 488-nm, single line laser for fluorophore excitation. The optical slice thickness is 7 μm, the lateral resolution 0.7 μm. The range of the z-axis is 0 - 250 μm below the tissue surface. Imaging was performed using different fluorescent staining protocols; 5-carboxyfluorescein-labeled octreotate was synthesized for targeted molecular imaging.

Results: Cellular and subcellular details of the gastrointestinal tract could be visualized in vivo at high resolution. Confocal real-time microscopy allowed in-vivo identification of tumor vessels and liver metastases, as well as diagnosis of focal hepatic inflammation, necrosis, and associated perfusion anomalies. Somatostatin-receptor targeting permitted in-vivo molecular staining of AR42-J-induced carcinoma and pancreatic islet cells.

Conclusions: Confocal mini-microscopy allows rapid in-vivo molecular and subsurface imaging of normal and pathological tissue in the gastrointestinal tract at high resolution. Because this technology is applicable to humans, it might impact on future in-vivo microsocpic and molecular diagnosis of diseases such as cancer and inflammation.

References

• 1 Stephens D, Allan V J. Light microscopy techniques for in vivo imaging.  Science. 2003;  300 82-86
  CrossrefPubMedGoogle Scholar
• 2 Wright S J, Wright D J. Introduction to confocal microscopy.  Methods Cell Biol. 2002;  70 1-85
  PubMedGoogle Scholar
• 3 Dacosta R S, Wilson B C, Marcon M E. New optical technologies for earlier endoscopic diagnosis of premalignant gastrointestinal lesions.  J Gastroenterol Hepatol. 2002;  17 S85-S104
  CrossrefPubMedGoogle Scholar
• 4 Sokolov K, Aaron J, Hsu B. et al . Optical systems for in vivo molecular imaging of cancer.  Technol Cancer Res Treat. 2003;  2 491-504
  PubMedGoogle Scholar
• 5 Flusberg B A, Cocker E D, Piyawattanametha W. et al . Fiber-optic fluorescence imaging.  Nat Methods. 2005;  2 941-950
  CrossrefPubMedGoogle Scholar
• 6 Kiesslich R, Burg J, Vieth M. et al . Confocal laser endoscopy for diagnosing intraepithelial neoplasias and colorectal cancer in vivo.  Gastroenterology. 2004;  127 706-713
  CrossrefPubMedGoogle Scholar
• 7 Nathanson M H. Confocal colonoscopy: more than skin deep.  Gastroenterology. 2004;  127 987-989
  CrossrefPubMedGoogle Scholar
• 8 Schirrmacher E, Schirrmacher R, Beck C. et al . Synthesis of a Tyr3-octreotate conjugated closo-carborane (HC2B10H10): a potential compound for boron neutron capture therapy.  Tetrahedron Letters. 2003;  44 9143-9145
  CrossrefPubMedGoogle Scholar
• 9 Weber P, Bader J, Folkers G. et al . A fast and inexpensive method for N-terminal fluorescein-labeling of peptides.  Bioorg Med Chem Lett. 1998;  8 597-600
  CrossrefPubMedGoogle Scholar
• 10 Anlauf M, Perren A, Meyer C L. et al . Precursor lesions in patients with multiple endocrine neoplasia type 1-associated duodenal gastrinomas.  Gastroenterology. 2005;  128 1187-1198
  CrossrefPubMedGoogle Scholar
• 11 Becker C, Fantini M C, Schramm C. et al . TGF-beta suppresses tumor progression in colon cancer by inhibition of IL-6 trans-signaling.  Immunity. 2004;  21 491-501
  CrossrefPubMedGoogle Scholar
• 12 Perez J, Viollet C, Doublier S. et al . Somatostatin binds to murine macrophages through two distinct subsets of receptors.  J Neuroimmunol. 2003;  138 38-44
  CrossrefPubMedGoogle Scholar
• 13 Sakashita M, Inoue H, Kashida H. et al . Virtual histology of colorectal lesions using laser-scanning confocal microscopy.  Endoscopy. 2003;  35 1033-1038
  Article in Thieme ConnectPubMedGoogle Scholar
• 14 Inoue H, Igari T, Nishikage T. et al . A novel method of virtual histopathology using laser-scanning confocal microscopy in-vitro with untreated fresh specimens from the gastrointestinal mucosa.  Endoscopy. 2000;  32 439-443
  Article in Thieme ConnectPubMedGoogle Scholar
• 15 Keller R, Winde G, Terpe H J. et al . Fluorescence endoscopy using a fluorescein-labelled monoclonal antibody against carcinoembryonic antigen in patients with colorectal carcinoma and adenoma.  Endoscopy. 2002;  34 801-807
  Article in Thieme ConnectPubMedGoogle Scholar
• 16 Weissleder R, Tung C H, Mahmood U. et al . In vivo imaging of tumors with protease-activated near-infrared fluorescent probes.  Nat Biotech. 1999;  17 375-378
  CrossrefPubMedGoogle Scholar
• 17 Kelly K, Alencar H, Funovics M. et al . Detection of invasive colon cancer using a novel, targeted, library-derived fluorescent peptide.  Cancer Res. 2004;  64 6247-6251
  CrossrefPubMedGoogle Scholar
• 18 Hsu E R, Anslyn E V, Dharmawardhane S. et al . A far-red fluorescent contrast agent to image epidermal growth factor receptor expression.  Photochem Photobiol. 2004;  79 272-279
  CrossrefPubMedGoogle Scholar

1 * The first two authors contributed equally to this paper.

2 # These authors share senior authorship.

M. Goetz, MD

Medizinische Klinik I

Johannes Gutenberg-Universität Mainz

Langenbeckstr. 1

55131 Mainz

Germany

Fax: +49-6131-17-6416

Email: mgoetz@mail.uni-mainz.de