[Physical principles of optical biopsy].

The purpose of this presentation is to explain the physical principles underlying the three main methods used to obtain images of living tissues at the cellular scale. In confocal microscopy, the tissue of interest is illuminated and scanned through a confocal aperture; a lateral resolution close to 1 microm can be obtained with high numerical aperture. Multiphoton microscopy uses a high-power short-pulse laser with instantaneous irradiance sufficient to excite fluorescence in a very small focal volume. The concentration of natural tissue fluorophores is too low to obtain an adequate signal, so exogenous fluorophores have to be added, either locally or through the body. These fluorophores can be conjugated to a variety of biomolecules that target specific disease processes, thereby increasing diagnostic specificity. Finally, OCT (optical coherence tomography) provides high-resolution images of entire tissue volumes by using a broadband source and an interferometric configuration; the depth of field and lateral resolution are both on the micrometer scale. These methods allow images to be obtained at the cellular level, but image contrast and stability require specific adjustment for their significance to be established with respect to conventional biopsy methods.