Multiphoton imaging of cardiovascular structures

Near infrared (NIR) femtosecond laser imaging systems represent a novel and very promising diagnostic technology for non-invasive cross-sectional analysis of living biological tissues. In this study 3D multiphoton imaging with submicron resolution has been performed for non-invasive analysis of living native and tissue-engineered (TE) heart valves and blood vessels. High-resolution autofluorescence and second harmonic generation (SHG) images of collagenous structures and elastic fibers were demonstrated using multiphoton excitation at two different wavelengths. Non-invasive optical sections have been obtained without the need of staining or embedding. The quality of the resulting three-dimensional images allowed exact differentiation between collagenous structures and elastic fibers. These experimental results are very encouraging for NIR femtosecond laser scanning microscopy as a useful tool for future non-destructive monitoring and characterization of vital and intact TE cardiovascular structures.

[1]  F J Schoen,et al.  Aortic valve structure-function correlations: role of elastic fibers no longer a stretch of the imagination. , 1997, The Journal of heart valve disease.

[2]  Russell Hk,et al.  A modification of Movat's pentachrome stain. , 1972 .

[3]  U A Stock,et al.  Tissue engineering of heart valves -- current aspects. , 2002, The Thoracic and cardiovascular surgeon.

[4]  Iris Riemann,et al.  High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution. , 2003, Journal of biomedical optics.

[5]  K J Halbhuber,et al.  Complete dynamic repopulation of decellularized heart valves by application of defined physical signals-an in vitro study. , 2003, Cardiovascular research.

[6]  Hartmut Oehring,et al.  Cytochemical demonstration of expression and distribution of non-glycosylated human lysosomal cathepsin S in HEK 293 cells. , 2002, Cellular and molecular biology.

[7]  Guy Cox,et al.  3-dimensional imaging of collagen using second harmonic generation. , 2003, Journal of structural biology.

[8]  B. Tromberg,et al.  Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[9]  K. Svoboda,et al.  Two-photon imaging in living brain slices. , 1999, Methods.

[10]  K J Halbhuber,et al.  Impact of decellularization of xenogeneic tissue on extracellular matrix integrity for tissue engineering of heart valves. , 2003, Journal of structural biology.

[11]  Paul J Campagnola,et al.  Second harmonic generation imaging of endogenous structural proteins. , 2003, Methods.

[12]  William A Mohler,et al.  Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues. , 2002, Biophysical journal.

[13]  S. Akhtar,et al.  Immunolocalization of elastin, collagen type I and type III, fibronectin, and vitronectin in extracellular matrix components of normal and myxomatous mitral heart valve chordae tendineae. , 1999, Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology.

[14]  K König,et al.  Technical advance: near-infrared femtosecond laser pulses as a novel non-invasive means for dye-permeation and 3D imaging of localised dye-coupling in the Arabidopsis root meristem. , 1999, The Plant journal : for cell and molecular biology.

[15]  P. So,et al.  Two-Photon deep tissue ex vivo imaging of mouse dermal and subcutaneous structures. , 1998, Optics express.

[16]  H. Schneckenburger,et al.  Laser-induced autofluorescence for medical diagnosis , 1994, Journal of Fluorescence.

[17]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.