C. Depeursinge

Photodetection and photodynamic therapy of "early" squamous cell carcinomas of the pharynx, oesophagus and tracheo-bronchial tree

The efficacy of photodynamic therapy (PDT) alone was evaluated on 41 ‘early’ squamous cell carcinomas of the pharynx (10), oesophagus (15) and tracheo-bronchial tree (16). All lesions but two were synchronous second primaries in ENT-patients suffering from a more extensive cancer, governing the overall oncological prognosis.Photofrin I (3 mg/kg) or Photofrin II (2 mg/kg) were injected 72 h prior to the red light irradiation, supplied by an argon pumped dye laser. A diffusing cylinder was used to obtain a homogeneous light distribution at the tumour site (60 J to 150 J/cm2). In the oesophagus and bronchi, the results are good for cancers staged in situ or microinvasive at endoscopy (two recurrencies for 23 lesions treated). For more advanced cancers (submucosal in the oesophagus or invading the bronchial cartilage), the results are less satisfactory (three recurrencies for eight lesions treated). In the pharynx where light dosimetry is more difficult, the rate of recurrencies is higher (3/10 lesions treated). In the bronchi (one case) and oesophagus (one case), the longest disease-free survival is now 5 years.The irradiation of a non-cancerous zone of normal buccal mucosa on 25 patients having received HPD showed necrosis in all cases with light doses as low as 50mW/cm2 for 20 min (60 J cm−2), even with Photofrin II.We encountered six complications (three cicatricial stenosis, two fistulae, one severe sunburn), most of them resulting from the lack of selectivity of HPD. According to these experiments, PDT is efficient at destroying early squamous cell carcinomas in the pharynx, oesophagus and bronchi, but the tumour selectivity of HPD is poor in the digestive tract lined with squamous cell epithelium. The only hope for the future lies in the synthesis of a more selective and more stable photosensitizer. This discussion reviews possible directions of research for the development of new dyes (cationic dyes, dyes attached to monoclonal antibodies, etc), for PDT and hyperthermia, for photodetection of early cancers using a fluoro-endoscope, and finally, for tumour depth profiling in hollow organs using lasers of different wavelengths.

Multiplexed 3D imaging using wavelength encoded spectral interferometry: a proof of principle

Based on spectral interferometry and wavelength multiplexing, we demonstrate that a combination of these methods could lead to a 3D imaging system. This system has the advantage of a one-dimensional geometry transmission channel and a limited scanning need. This kind of imaging system could, of course, be of a great interest in small size endoscope with 3D imaging capability and a small channel cross-section (less than 1 mm). Theoretical analysis as well as experimental proof of principle is presented in the following paper.

Simulation of time-resolved breast transillumination

A Monte Carlo simulation has been developed to predict the quality of time-resolved images of the breast by transillumination. The smallest diameter of a detectable carcinoma located in the breast has been computed. The simulation suggests that time-resolved imaging of the breast is possible and invaluable in the near infra-red (NIR) by transillumination. The enhancement of the transfer function by the introduction of time-resolved detection is limited by the contribution of noise at short integration times. The estimated diameter of the smallest detectable sphere is derived from the image quality index (IQI) theory and its value is around 4 mm. The simulated images of an absorbing sphere (approximating the carcinoma) within a homogeneous medium (approximating the surrounding tissue) show a significant improvement of the image with short integration time.

Analysis of cellular structure and dynamics with digital holographic microscopy

Digital holographic microscopy (DHM) is a technique that allows obtaining, from a single recorded hologram, quantitative phase image of living cell with interferometric accuracy. Specifically, the optical phase shift induced by the specimen on the transmitted wave front can be regarded as a powerful endogenous contrast agent, depending on both the thickness and the refractive index of the sample. We discuss some approaches allowing to directly obtain separate measurements of the thickness and the refractive index (RI) of a given living cell from the phase signal.

In vivo measurement of dye concentration using an evanescent-wave optical sensor.

A miniaturised evanescent-wave optical sensor is proposed for in vivo measurement of dye concentrations. It enables a continuous monitoring of the optical-dye attenuation or fluorescence spectra between 380 and 650 nm. The sensor is constructed with polished fibres: the cladding of a single-mode fibre is removed by longitudinal polishing. The proximity of the core to the medium favours penetration of the evanescent part of the modal field into the bio fluid. The dimensions of the probe permit several potential applications: for example, insertion into hypodermic needles for spectroscopic analysis of tissues and blood. In the paper, a gastro-enterologic application of the sensor introduced into a catheter is reported. In vivo tests demonstrate the feasibility of quantitative measurement of dye clearance in the gastro-oesophageal tract.

Red blood cell structure and dynamics explored with digital holographic microscopy

Digital holographic microscopy (DHM) is a technique that allows obtaining, from a single recorded hologram, quantitative phase image of living cell with interferometric accuracy. Specifically the optical phase shift induced by the specimen on the transmitted wave front can be regarded as a powerful endogenous contrast agent, depending on both the thickness and the refractive index of the sample. Thanks to a decoupling procedure cell thickness and intracellular refractive index can be measured separately. Consequently, Mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), two highly relevant clinical parameters, have been measured non-invasively at a single cell level. The DHM nanometric axial and microsecond temporal sensitivities have permitted to measure the red blood cell membrane fluctuations (CMF) on the whole cell surface.

Wavelength Multiplexed Spectral Interferometry for Endoscopic Topographic Imaging

We present an imaging system which could be used for endoscopic topography. Indeed it allows for two dimensional information transmission through a unidimensional imaging channel which is a monomode optical fiber. The principle is the coupling of wavelength multiplexing and spectral interferometry and a special configuration renders this system dispersion self compensated what enables a high signal stability. Principles will be presented as well as results and limits of the system

Erythrocytes analysis with a Digital Holographic Microscope

Digital holographic microscopy (DHM) is a technique that allows obtaining, from a single recorded hologram, quantitative phase image of living cell with interferometric accuracy (Marquet et al., 2005). Specifically the optical phase shift induced by the specimen on the transmitted wave front can be regarded as a powerful endogenous contrast agent, depending on both the thickness and the refractive index of the sample. We have recently proposed (Rappaz et al., 2005) a new and efficient decoupling procedure allowing to directly obtain separate measurements of the thickness and the integral refractive index of a given living cell. Consequently, it has been possible, for the first time to our knowledge, to accurately measure (with a precision of 0.0003) the mean refractive index of living erythrocytes.. On the other hand, the cellular thickness measurements allow to calculate the volume and shape of erythrocytes. In addition, DHM, thanks to its subwavelength phase shift measurements, was found to yield an efficient tool to assess erythrocyte cell membrane fluctuations (ECMF). Typically, ECMF characterized by an amplitude within the range of 45 nm were observed.

Simultaneous Objective Measurements Of Dose And Image Quality In Mammography

The performance of a radiological system can be evaluated on the one hand by an objective determination of the quality of the produced image and, on the other hand, by the dose delivered to the patient. In order to measure these two factors in a single exposure a Kodak breast phantom has been modified so as to simulate the breast absorption. The dose distribution is measured with thermoluminescent detectors. By consideration of a theoretical model of the X-ray imaging in mammography, a single quality factor is computed from the contrast, the spatial resolution and the noise measured on the phantom image. We present results obtained in various working conditions, i.e. variable X-ray tube voltages, use of different screen-film combinations, use of a grid.

Cellular Dynamics Revealed by Digital Holographic Microscopy

Digital holographic microscopy (DHM) is a new optical method that provides, without the use of any contrast agent, real-time, three-dimensional images of transparent living cells, with an axial sensitivity of a few tens of nanometers. They result from the hologram numerical reconstruction process, which permits a sub wavelength calculation of the phase shift, produced on the transmitted wave front, by the optically probed cells, namely the quantitative phase signal (QPS). Specifically, in addition to measurements of cellular surface morphometry and intracellular refractive index (RI), various biophysical cellular parameters including dry mass, absolute volume, membrane fluctuations at the nanoscale and biomechanical properties, transmembrane water permeability as swell as current, can be derived from the QPS. This article presents how quantitative phase DHM (QP-DHM) can explored cell dynamics at the nanoscale with a special attention to both the study of neuronal dynamics and the optical resolution of local neuronal network.