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Biomedical subjects

Marcel Leutenegger

Publications and source records attributed to Marcel Leutenegger.

4 recordsLinked to original sources

Prism-based multicolor fluorescence correlation spectrometer.

We report the design and application of a prism-based detection system for fluorescence (cross) correlation spectroscopy. The system utilizes a single laser wavelength for the simultaneous excitation of several dyes of different emission spectra. Fluorescence light is spectrally separated with a prismatic setup, and wavelengths are selected by scanning a fiber-coupled avalanche photodiode across the image spots. Multicolor autocorrelations are demonstrated with standard and tandem dyes, and fluorescence cross-correlation measurements of biotinylated nanocontainers and streptavidin are presented. This spectrometer offers high optical stability and no focal volume mismatch for the multicolor detection of molecular dynamics and interactions, with single-molecule sensitivity.

Equipment Design↗

Multiple scattering in optical coherence tomography. I. Investigation and modeling.

We present a new model of optical coherence tomography (OCT) taking into account multiple scattering. A theoretical analysis and experimental investigation reveals that in OCT, despite multiple scattering, the field backscattered from the sample is generally spatially coherent and that the resulting interference signal with the reference field is stationary relative to measurement time. On the basis of this result, we model an OCT signal as a sum of spatially coherent fields with random-phase arguments--constant during measurement time--caused by multiple scattering. We calculate the mean of such a random signal from classical results of statistical optics and a Monte Carlo simulation. OCT signals predicted by our model are in very good agreement with a depth scan measurement of a sample consisting of a mirror covered with an aqueous suspension of microspheres. We discuss other comprehensive OCT models based on the extended Huygens-Fresnel principle, which rest on the assumption of partially coherent interfering fields.

Algorithms↗

Multiple scattering in optical coherence tomography. II. Experimental and theoretical investigation of cross talk in wide-field optical coherence tomography.

We present a comprehensive study of multiple-scattering effects in wide-field optical coherence tomography (OCT) realized with spatially coherent illumination. Imaging a sample made of a cleaved mirror embedded in an aqueous suspension of microspheres revealed that, despite temporal coherence gating, multiple scattering can induce significant coherent optical cross talk. The latter is a serious limitation to the method, since it prevents shot-noise-limited detection and diffraction-limited imaging in scattering samples. We investigate the dependence of cross talk on important system design parameters, as well as on some relevant sample properties. The agreement between theoretical and experimental results for the wide range of parameters investigated was very good, in both the lateral and the axial dimensions. This further confirms the validity of the model developed in our companion paper [J. Opt. Soc. Am. A 22, 1369-1379 (2005)].

Algorithms↗

Dual-color total internal reflection fluorescence cross-correlation spectroscopy.

We present the development and first application of a novel dual-color total internal reflection (TIR) fluorescence system for single-molecule coincidence analysis and fluorescence cross-correlation spectroscopy (FCCS). As a performance analysis, we measured a synthetic DNA-binding assay, demonstrating this dual-color TIR-FCCS approach to be a suitable method for measuring coincidence assays such as biochemical binding, fusion, or signal transduction at solid/liquid interfaces. Due to the very high numerical aperture of the epi-illumination configuration, our setup provides a very high fluorescence collection efficiency resulting in a two- to three-fold increase in molecular brightness compared to conventional confocal FCCS. Further improvements have been achieved through global analysis of the spectroscopic data.

DNA↗