A multispectral FLIM tomograph for in-vivo imaging of skin cancer

To aid the in vivo diagnosis of skin lesions, we present the design and implementation of a 4 channel FLIM detector and a hyperspectral imaging detector into a clinically licensed commercial two-photon tomograph. We have also implemented image segmentation algorithms to facilitate the automated processing of the large volumes of data produced. The first detector is based on multispectral time correlated single photon counting, providing four channel fluorescence lifetime images. The second detector is a prism-based CCD hyperspectral imager. These detectors provide the capability to extract the relative content and state of autofluorescence compounds present in biological tissue.

[1]  Michael Zuker,et al.  Delta function convolution method (DFCM) for fluorescence decay experiments , 1985 .

[2]  Dong Li,et al.  Autofluorescence of epithelial tissue: single-photon versus two-photon excitation. , 2008, Journal of biomedical optics.

[3]  Jens Eickhoff,et al.  In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia. , 2007, Journal of biomedical optics.

[4]  P. Carli,et al.  Multidimensional non-linear laser imaging of Basal Cell Carcinoma. , 2007, Optics express.

[5]  Yong-Gu Lee,et al.  Simulation of an oil immersion objective lens: a simplified ray-optics model considering Abbe's sine condition. , 2008, Optics express.

[6]  A. Bergmann,et al.  Multispectral fluorescence lifetime imaging by TCSPC , 2007, Microscopy research and technique.

[7]  Hans C Gerritsen,et al.  Design and implementation of a sensitive high-resolution nonlinear spectral imaging microscope. , 2008, Journal of biomedical optics.

[8]  Karsten König,et al.  Spectral fluorescence lifetime detection and selective melanin imaging by multiphoton laser tomography for melanoma diagnosis , 2009, Experimental dermatology.

[9]  Watt W Webb,et al.  Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein. , 2002, Biophysical journal.

[10]  B. Wilson,et al.  In Vivo Fluorescence Spectroscopy and Imaging for Oncological Applications , 1998, Photochemistry and photobiology.

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

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

[13]  W. Webb,et al.  Conformational Dependence of Intracellular NADH on Metabolic State Revealed by Associated Fluorescence Anisotropy*♦ , 2005, Journal of Biological Chemistry.

[14]  L. Gaboury,et al.  Steady state and time-resolved fluorescence properties of metastatic and non-metastatic malignant cells from different species. , 1995, Journal of photochemistry and photobiology. B, Biology.

[15]  Yuriy Alexandrov,et al.  Angiogenesis: an improved in vitro biological system and automated image-based workflow to aid identification and characterization of angiogenesis and angiogenic modulators. , 2008, Assay and drug development technologies.

[16]  Christophe Odin,et al.  Non-descanned versus descanned epifluorescence collection in two-photon microscopy: Experiments and Monte Carlo simulations , 2008 .

[17]  Klaus Hoffmann,et al.  Fluorescence Studies of Melanin by Stepwise Two-Photon Femtosecond Laser Excitation , 2000, Journal of Fluorescence.

[18]  Wei Zheng,et al.  Sensing cell metabolism by time-resolved autofluorescence. , 2006, Optics letters.

[19]  H. S. de Bruijn,et al.  In vivo nonlinear spectral imaging in mouse skin. , 2006, Optics express.

[20]  Hans C Gerritsen,et al.  Spectrally resolved multiphoton imaging of in vivo and excised mouse skin tissues. , 2007, Biophysical journal.