Nicolas Verrier

Digital in-line holography in thick optical systems: application to visualization in pipes

We apply digital in-line holography to image opaque objects through a thick plano-concave pipe. Opaque fibers and opaque particles are considered. Analytical expression of the intensity distribution in the CCD sensor plane is derived using a generalized Fresnel transform. The proposed model has the ability to deal with various pipe shapes and thicknesses and compensates for the lack of versatility of classical digital in-line holography models. Holograms obtained with a 12 mm thick plano-concave pipe are then reconstructed using a fractional Fourier transform. This method allows us to get rid of astigmatism. Numerical and experimental results are presented.

Micropipe flow visualization using digital in-line holographic microscopy

Digital in-line holography is used to visualize particle motion within a cylindrical micropipe. Analytical expression of the intensity distribution recorded in the CCD sensor plane is derived using the generalized Huygens-Fresnel integral associated with the ABCD matrices formalism. Holograms obtained in a 100microm in diameter micropipe are then reconstructed using fractional Fourier transformation. Astigmatism brought by the cylindrical micropipe is finally used to select a three dimensional region of interest in the microflow and thus to improve axial localization of objects located within a micropipe. Experimental results are presented and a short movie showing particle motion within a micropipe is given.

Laser Doppler holographic microscopy in transmission: application to fish embryo imaging

We have extended Laser Doppler holographic microscopy to transmission geometry. The technique is validated with living fish embryos imaged by a modified upright bio-microcope. By varying the frequency of the holographic reference beam, and the combination of frames used to calculate the hologram, multimodal imaging has been performed. Doppler images of the blood vessels for different Doppler shifts, images where the flow direction is coded in RGB colors or movies showing blood cells individual motion have been obtained as well. The ability to select the Fourier space zone that is used to calculate the signal, makes the method quantitative.

Pixel super-resolution in digital holography by regularized reconstruction

In-line digital holography (DH) and lensless microscopy are 3D imaging techniques used to reconstruct the volume of micro-objects in many fields. However, their performances are limited by the pixel size of the sensor. Recently, various pixel super-resolution algorithms for digital holography have been proposed. A hologram with improved resolution was produced from a stack of laterally shifted holograms, resulting in better resolved reconstruction than a single low-resolution hologram. Algorithms for super-resolved reconstructions based on inverse problems approaches have already been shown to improve the 3D reconstruction of opaque spheres. Maximum a posteriori (MAP) approaches have also been shown capable of reconstructing the object field more accurately and more efficiently and to extend the usual field-of-view. Here we propose an inverse problem formulation for DH pixel super-resolution and an algorithm that alternates registration and reconstruction steps. The method is described in detail and used to reconstruct synthetic and experimental holograms of sparse 2D objects. We show that our approach improves both the shift estimation and reconstruction quality. Moreover, the reconstructed field-of-view can be expanded by up to a factor 3, thus making it possible to multiply the analyzed area 9 fold.

Phase-resolved heterodyne holographic vibrometry with a strobe local oscillator

We report a demonstration of phase-resolved vibrometry, in which out-of-plane sinusoidal motion is assessed by heterodyne holography. In heterodyne holography, the beam in the reference channel is an optical local oscillator (LO). It is frequency-shifted with respect to the illumination beam to enable frequency conversion within the sensor bandwidth. The proposed scheme introduces a strobe LO, where the reference beam is frequency-shifted and modulated in amplitude, to alleviate the issue of phase retrieval. The strobe LO is both tuned around the first optical modulation sideband at the vibration frequency, and modulated in amplitude to freeze selected mechanical vibration states sequentially. The phase map of the vibration can then be derived from the demodulation of successive vibration states.

Holographic microscopy reconstruction in both object and image half spaces with undistorted 3D grid

We propose a holographic microscopy reconstruction method that propagates the hologram in the object half-space in the vicinity of the object. The calibration yields reconstructions with an undistorted reconstruction grid, i.e., with orthogonal x, y, and z axes and constant pixel pitch. The method is validated with a USAF target imaged by a ×60 microscope objective (MO), whose holograms are recorded and reconstructed for different USAF locations along the longitudinal axis: -75 to +75  μm. Since the reconstruction numerical phase mask, the reference phase curvature, and the MO form an afocal device, the reconstruction can be interpreted as occurring equivalently in the object or image half-space.

Vibration of low amplitude imaged in amplitude and phase by sideband versus carrier correlation digital holography

Sideband holography can be used to get field images (E0 and E1) of a vibrating object for both the carrier (E0) and the sideband (E1) frequency with respect to vibration. Here we propose to record E0 and E1 sequentially and to image the product E1E0* or the correlation 〈E1E0*〉. We show that these quantities are insensitive to the phase related to the object roughness and directly reflect the phase of the mechanical motion. The signal to noise can be improved by averaging E1E0* over a neighbor pixel, yielding 〈E1E0*〉. Experimental validation is made with a vibrating cube of wood and a clarinet reed. At 2 kHz, vibrations of amplitude down to 0.01 nm are detected.

Holographic microscopy reconstruction in both object and image half-spaces with an undistorted three-dimensional grid

We propose a holographic microscopy reconstruction method that propagates the hologram in the object half-space in the vicinity of the object. The calibration yields reconstructions with an undistorted reconstruction grid, i.e., with orthogonal x, y, and z axes and constant pixel pitch. The method is validated with a USAF target imaged by a ×60 microscope objective (MO), whose holograms are recorded and reconstructed for different USAF locations along the longitudinal axis: -75 to +75  μm. Since the reconstruction numerical phase mask, the reference phase curvature, and the MO form an afocal device, the reconstruction can be interpreted as occurring equivalently in the object or image half-space.

High numerical aperture holographic microscopy reconstruction with extended z range.

A holographic microscopy reconstruction method compatible with a high numerical aperture microscope objective (MO) up to NA=1.4 is proposed. After off-axis and reference field curvature corrections, and after selection of the +1 grating order holographic image, a phase mask that transforms the optical elements of the holographic setup into an afocal device is applied in the camera plane. The reconstruction is then made by the angular spectrum method. The field is first propagated in the image half-space from the camera to the afocal image of the MO optimal plane (the plane for which the MO has been designed) by using a quadratic kernel. The field is then propagated from the MO optimal plane to the object with the exact kernel. Calibration of the reconstruction is made by imaging a calibrated object such as a USAF resolution target for different positions along z. Once the calibration is done, the reconstruction can be made with an object located in any plane z. The reconstruction method has been validated experimentally with a USAF target imaged with a NA=1.4 microscope objective. Near-optimal resolution is obtained over an extended range (±50  μm) of z locations.

Self-calibration for lensless color microscopy

Lensless color microscopy (also called in-line digital color holography) is a recent quantitative 3D imaging method used in several areas including biomedical imaging and microfluidics. By targeting cost-effective and compact designs, the wavelength of the low-end sources used is known only imprecisely, in particular because of their dependence on temperature and power supply voltage. This imprecision is the source of biases during the reconstruction step. An additional source of error is the crosstalk phenomenon, i.e., the mixture in color sensors of signals originating from different color channels. We propose to use a parametric inverse problem approach to achieve self-calibration of a digital color holographic setup. This process provides an estimation of the central wavelengths and crosstalk. We show that taking the crosstalk phenomenon into account in the reconstruction step improves its accuracy.