In-vivo tissue imaging using a compact mobile nonlinear microscope

Year: 2010

Authors: Cicchi R., Kapsokalyvas D., Stambouli D., De Giorgi V., Massi D., Lotti T., Pavone FS.

Autors Affiliation: European Laboratory for Non-linear Spectroscopy (LENS), University of Florence, 1 Via Nello Carrara, 50019 Sesto Fiorentino, Italy; Department of Dermatological Sciences, University of Florence, Florence, Italy; Department of Human Pathology and Oncology, University of Florence, Florence, Italy

Abstract: We have built a compact flexible non-linear microscope equipped with a combination of different non-linear laser imaging techniques including two-photon fluorescence, second-harmonic generation, fluorescence lifetime imaging microscopy, and multispectral two-photon emission detection. The system is composed of a microscope head, containing both scanning and detection system, as well as the electronic and electro-mechanical devices, optically relayed to the laser source with a seven-mirror articulated arm. The particular mirror positioning inside the arm allows to move the microscope head maintaining the optical alignment of the system. The microscope head is composed by two ErGaAl anodized boards, one for laser scanning and the other for signal detection. System performances were characterized by means of point spread function and instrument response function measurements as well as by spatial, temporal, and spectral calibration. The instrument, offering high spatial (up to 300 nm) and temporal (up to 300 ps) resolution, was tested on in-vivo skin imaging of both cellular epidermis and connective dermis. Lifetime and spectral features of fluorescence were used for differentiating epidermal layers by means of fluorescence lifetime and for scoring skin ageing through spectral detection of both second-harmonic and two-photon fluorescence.

Conference title: Photonics West – BIOS 2010
Place: San Francisco, US

KeyWords: Detection system; Electro-mechanical; Epidermal layers; Fluorescence lifetime imaging microscopy; Fluorescence lifetimes; In-vivo; Instrument response functions; Laser scanning; Laser sources; Lifetime imaging; Multi-photon microscopy; Multi-spectral; Multispectral imaging; Non-linear; Optical alignments; Optical biopsies; Point-Spread function; Second harmonic generation; Skin imaging; Spectral calibration; Spectral detection; Spectral feature; Two photon fluorescence; Two-photon emission, Biopsy; Electromechanical devices; Fluorescence; Imaging techniques; Microscopes; Optical transfer function; Photons, Mirrors
DOI: 10.1117/12.841752

Citations: 14
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