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GJ Brakenhoff

Publications and source records attributed to GJ Brakenhoff.

4 recordsLinked to original sources

CARS microscopy with folded BoxCARS phasematching

Three-dimensional microscopy based on coherent anti-Stokes Raman scattering (CARS) is a powerful new imaging technique, in which the contrast arises from molecular vibrations. Based on a simple numerical model, it is shown how the CARS interaction volume depends on the focusing parameters and the type of phasematching used. Collinear phasematching yields an ellipsoidal interaction volume, with lateral dimensions that readily cause vignetting of the CARS signal emission at the collection microscope objective. A folded BoxCARS phasematching geometry, on the other hand, results in an almost cylindrical interaction volume - at the cost of a reduced resolution, for which the possible vignetting of the CARS emission is much reduced. In addition, this type of phasematching provides spatial separation of the signal from the input laser beams, permitting simple signal detection of low frequency vibrational modes. Calculations show that when CARS is performed in a microscopic volume, the phasematching restraint on tuning over the vibrational band is strongly relaxed. A first example of CARS imaging using a folded BoxCARS imaging geometry is shown.

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Fluorescence photobleaching-based image standardization for fluorescence microscopy

A method is presented for the standardization of images acquired with fluorescence microscopy, based on the knowledge of spatial distributions proportional to the microscope's absolute excitation intensity and fluorescence detection efficiency distributions over the image field. These distributions are determined using a thin fluorescent test layer, employed under practically mono-exponential photobleaching conditions. It is demonstrated that these distributions can be used for (i) the quantitative evaluation of differences between both the excitation intensity and the fluorescence detection efficiency of different fluorescence microscopes and (ii) the standardization of images acquired with different microscopes, permitting the deduction of quantitative relationships between images obtained under different imaging conditions.

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Dispersion pre-compensation of 15 femtosecond optical pulses for high-numerical-aperture objectives.

The excitation efficiency in two-photon absorption (TPA) microscopy depends strongly - owing to the square dependence of the TPA fluorescence on the excitation intensity - on the temporal width of the excitation pulse. Because of their inherently large frequency bandwidth, ultrashort optical pulses tend to broaden substantially because of dispersion from propagation through the dispersive elements in the microscope. In this paper, the dispersion characteristics of a wide range of microscope objectives are investigated. It is shown that the induced dispersion can be pre-compensated in all cases for pulses as short as 15 fs. Because of the excellent agreement between the results from theoretical modelling and the experimental data, predictions of the possibility of dispersion control for microscope objectives in general, as well as for even shorter pulses, can be inferred. Since for TPA imaging the background due to single photon absorption processes and scattering is independent of the pulse width, proper dispersion pre-compensation - which minimizes the pulse duration at the focal point and hence maximizes the excitation efficiency - provides optimal image contrast in TPA microscopy.

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3D microscopy of transparent objects using third-harmonic generation.

It is demonstrated that third-harmonic generation (THG) near interfaces in the refractive index or the third-order nonlinear susceptibility (chi(3)) permits three-dimensional imaging of transparent objects. The nonlinear dependence of THG on the excitation power provides inherent optical sectioning. At the same time, the nonresonant nature of THG, in combination with the near-IR excitation wavelengths used (1-2 µm), render this technique potentially (biologically) nondamaging and nonbleaching. A specific property of THG imaging is its sensitivity to - and potential use for imaging of - the relative orientation of interfaces with respect to the axis of propagation of the excitation radiation.

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