Hyperspectral Interferometry Research Articles

Rapid, Highly Sensitive, and Label-Free Pathogen Assay System Using a Solid-Phase Self-Interference Recombinase Polymerase Amplification Chip and Hyperspectral Interferometry.

Recombinase polymerase amplification (RPA) is a useful pathogen identification method. Several label-free detection methods for RPA amplicons have been developed in recent years. However, these methods still lack sensitivity, specificity, efficiency, or simplicity. In this study, we propose a rapid, highly sensitive, and label-free pathogen assay system based on a solid-phase self-interference RPA chip (SiSA-chip) and hyperspectral interferometry. The SiSA-chips amplify and capture RPA amplicons on the chips, rather than irrelevant amplicons such as primer dimers, and the SiSA-chips are then analysed by hyperspectral interferometry. Optical length increases of SiSA-chips are used to demonstrate RPA detection results, with a limit of detection of 1.90 nm. This assay system can detect as few as six copies of the target 18S rRNA gene of Plasmodium falciparum within 20 min, with a good linear relationship between the detection results and the concentration of target genes (R2 = 0.9903). Single nucleotide polymorphism (SNP) genotyping of the dhfr gene of Plasmodium falciparum is also possible using the SiSA-chip, with as little as 1% of mutant gene distinguished from wild-type loci (m/wt). This system offers a high-efficiency (20 min), high-sensitivity (6 copies/reaction), high-specificity (1% m/wt), and low-cost (∼1/50 of fluorescence assays for RPA) diagnosis method for pathogen DNA identification. Therefore, this system is promising for fast identification of pathogens to help diagnose infectious diseases, including SNP genotyping.

Hyperspectral imaging for dynamic thin film interferometry

Dynamic thin film interferometry is a technique used to non-invasively characterize the thickness of thin liquid films that are evolving in both space and time. Recovering the underlying thickness from the captured interferograms, unconditionally and automatically is still an open problem. Here we report a compact setup employing a snapshot hyperspectral camera and the related algorithms for the automated determination of thickness profiles of dynamic thin liquid films. The proposed technique is shown to recover film thickness profiles to within 100 nm of accuracy as compared to those profiles reconstructed through the manual color matching process. Subsequently, we discuss the characteristics and advantages of hyperspectral interferometry including the increased robustness against imaging noise as well as the ability to perform thickness reconstruction without considering the absolute light intensity information.

Nine-hundred-channel single-shot surface roughness measurement using hyperspectral interferometry

A recently developed hyperspectral interferometer has been applied, for the first time, to the single-shot measurement of surface roughness. Traditional optical surface profiling techniques, such as coherence scanning interferometry (CSI) or focus variation microscopy, require long scan times and mechanical motion of the imaging objectives, making them vulnerable to environmental disturbances and thus inappropriate as an embedded metrology tool for production. Hyperspectral interferometry (HSI) answers these problems by capturing spatial and spectral information to reconstruct the surface profile, from 900 locations measured in parallel, in a single shot. We report here measurement results with both HSI and CSI from 12 individual roughness samples taken from Rubert roughness gauges covering the roughness range . Very good agreement was obtained for values in the range 3–30 µm, where the HSI values were mostly within a few per cent of (and at worst 24% away from) those measured by CSI . An alternative measure of roughness, based on averaging the widths of the Fourier transform peak of the individual HSI interference signals, is shown to be a statistically reliable measure of local roughness in the medium to high roughness regime, i.e. for values of 0.8 µm and above. The results thus demonstrate the technique’s potential for real-time surface quality inspection in manufacturing.

2500-Channel single-shot areal profilometer using hyperspectral interferometry with a pinhole array

Surface profilometry techniques such as coherent scanning interferometry or focus variation require long scan times and are thus vulnerable to environmental disturbance. Hyperspectral interferometry (HSI) overcomes the problem by recording all the spatial and spectral information necessary to reconstruct a 2D surface height map in a single shot. In this paper, we present a new HSI system that uses a pinhole array to provide the necessary gaps for the spectral information. It is capable of measuring 2500 independent points, twice the previous maximum number, with a maximum unambiguous depth range of ∼825 µm and a larger maximum surface tilt angle of 33.3 mrad. The use of phase information allows height to be measured to a precision of ∼6 nm, an order of magnitude improvement on previous HSI systems.

Deep UV dispersion and absorption spectroscopy of biomolecules.

Owing to the high precision and sensitivity of optical systems, there is an increasing demand for optical methods that quantitatively characterize the physical and chemical properties of biological samples. Information extracted from such quantitative methods, through phase and/or amplitude variations of light, can be crucial in the diagnosis, treatment and study of disease. In this work we apply a recently developed quantitative method, called ultraviolet hyperspectral interferometry (UHI), to characterize the dispersion and absorbing properties of various important biomolecules. Our system consists of (1) a broadband light source that spans from the deep-UV to the visible region of the spectrum, and (2) a Mach-Zehnder interferometer to gain access to complex optical properties. We apply this method to characterize (and tabulate) the dispersive and absorptive properties of hemoglobin, beta nicotinamide adenine dinucleotide (NAD), flavin adenine dinucleotide (FAD), elastin, collagen, cytochrome c, tryptophan and DNA. Our results shed new light on the complex properties of important biomolecules.

Single-shot areal profilometry using hyperspectral interferometry with a microlens array.

We describe a single-shot technique to measure areal profiles on optically smooth and stepped surfaces for applications where rapid data acquisition in non-cooperative environments is essential. It is based on hyperspectral interferometry (HSI), a technique in which the output of a white-light interferometer provides the input to a hyperspectral imaging system. Previous HSI implementations suffered from inefficient utilisation of the available pixels which limited the number of measured coordinates and/or unambiguous depth range. In the current paper a >20-fold increase in pixel utilisation is achieved through the use of a 2-D microlens array, that leads to a 35 × 35 channel system with an unambiguous depth range of 0.88 mm.

Label-Free Method Using a Weighted-Phase Algorithm To Quantitate Nanoscale Interactions between Molecules on DNA Microarrays.

White light interference is used as a label-free method to detect nanoscale changes on surfaces. However, the signal-to-noise ratio of the white light interference method is very low, thus resulting in inaccurate results. In this paper, we report a corrected label-free method based on hyperspectral interferometry to overcome the shortcoming of the white light interference method. A platform based on hyperspectral interferometry was established, and a DNA hybridization microarray was constructed to quantitate thickness variation of molecules on a solid surface. We used fluorescence resonance energy transfer (FRET) to validate the results of our method. Compared to conventional fluorescence-labeled method like FRET, our method has advantages because it does not require a fluorescent label and has a detection limit of 1.78 nm, a high accuracy, and wide detection range (5-64 bp).

Hyperspectral interferometry: Sizing microscale surface features in the pine bark beetle.

A new method of interferometry employing a Fabry-Perot etalon model was used to locate and size microscale features on the surface of the pine bark beetle. Oscillations in the reflected light spectrum, caused by self-interference of light reflecting from surfaces of foreleg setae and spores on the elytrum, were recorded using white light hyperspectral microscopy. By making the assumption that pairs of reflecting surfaces produce an etalon effect, the distance between surfaces could be determined from the oscillation frequency. Low frequencies of less than 0.08 nm(-1) were observed in the spectrum below 700 nm while higher frequencies generally occupied wavelengths from 600 to 850 nm. In many cases, two frequencies appeared separately or in combination across the spectrum. The etalon model gave a mean spore size of 3.04 ± 1.27 μm and a seta diameter of 5.44 ± 2.88 μm. The tapering near the setae tip was detected as a lowering of frequency. Spatial fringes were observed together with spectral oscillations from surfaces on the exoskeleton at higher magnification. These signals were consistent with embedded multi-layer reflecting surfaces. Possible applications for hyperspectral interferometry include medical imaging, detection of spore loads in insects and other fungal carriers, wafer surface and subsurface inspection, nanoscale materials, biological surface analysis, and spectroscopy calibration. This is, to our knowledge, the first report of oscillations directly observed by microscopy in the reflected light spectra from Coleoptera, and the first demonstration of broadband hyperspectral interferometry using microscopy that does not employ an internal interferometer.

Single-shot profilometry of rough surfaces using hyperspectral interferometry

The combination of white light interferometry with hyperspectral imaging ("hyperspectral interferometry") is a recently proposed technique for single-shot measurement of 3D surface profiles. We consider for the first time its application to speckled wavefronts from optically rough surfaces. The intensity versus wavenumber signal at each pixel provides unambiguous range information despite the speckle-induced random phase shifts. Experimental results with samples undergoing controlled rigid body translation demonstrate a measurement repeatability of 460 nm for a bandwidth of approximately 30 nm. Potential applications include roughness measurement and coordinate measurement machine probes where rapid data acquisition in noncooperative environments is essential.

Full field measurements and identification in Solid Mechanics Depth-resolved Phase Imaging

Traditional full-field interferometric techniques (speckle, moiré, holography etc) encode the surface deformation state of the object under test in the form of 2-D phase images. Over the past 10 years, a family of related techniques (Wavelength Scanning Interferometry, Phase Contrast Spectral Optical Coherence Tomography (OCT), Tilt Scanning Interferometry and Hyperspectral Interferometry) has emerged that allows one to measure the volume deformation state within weakly-scattering objects. The techniques can be thought of as combining the phase-sensing capabilities of Phase Shifting Interferometry and the depth-sensing capabilities of OCT. This paper provides an overview of the techniques, and describes a theoretical framework based on the Ewald sphere construction that allows key parameters such as depth resolution and displacement sensitivity to be calculated straightforwardly for any given optical geometry and wavelength scan range. Finally, the related issue of robust phase unwrapping of noisy 3-D wrapped phase volumes is also described.

Depth-resolved Imaging and Displacement Measurement Techniques Viewed as Linear Filtering Operations

The last 5 years have seen the emergence of a family of optical interferometric techniques that provide deformation measurements throughout three-dimensional (3-D) weakly scattering materials. They include wavelength scanning interferometry (WSI), tilt scanning interferometry (TSI), phase contrast spectral optical coherence tomography (PC SOCT) and hyperspectral interferometry (HSI) and can be thought of as a marriage between the phase sensing capabilities of Phase Shifting Interferometry and the depth-sensing capabilities of Optical Coherence Tomography. It was recently shown that some closely related 3-D optical imaging techniques can be treated as shift-invariant linear filtering operations. In this paper, we extend that work to include WSI, TSI, PC SOCT and HSI as spatial filtering operations and also relate the properties of their transfer functions in the spatial frequency domain to their spatial resolution and phase sensitivity, for depth-resolved displacement measurements.

Hyperspectral interferometry for single-shot absolute measurement of two-dimensional optical path distributions

We propose a method that we call hyperspectral interferometry (HSI) to resolve the 2π phase unwrapping problem in the analysis of interferograms recorded with a narrow-band light source. By using a broadband light source and a hyperspectral imaging system, a set of interferograms at different wave numbers are recorded simultaneously on a high resolution image sensor. These are then assembled to form a three-dimensional intensity distribution. By Fourier transformation along the wave number axis, an absolute optical path difference is obtained for each pixel independently of the other pixels in the field of view. As a result, interferograms with spatially distinct regions are analysed as easily as continuous ones. The approach is illustrated with a HSI system to measure 3D profiles of optically smooth or rough surfaces. Compared to existing profilometers able to measure absolute path differences, the single-shot nature of the approach provides greater immunity from environmental disturbance.

Hyperspectral interferometry for single-shot absolute measurement of 3-D shape and displacement fields

We propose a method that we call Hyperspectral Interferometry (HSI) to resolve the 2π phase unwrapping problem in the analysis of interferograms recorded with a narrow-band light source. By using a broad-band light source and hyperspectral imaging system, a set of interferograms at different wavenumbers are recorded simultaneously on a high resolution image sensor. These are then assembled to form a three-dimensional intensity distribution. By Fourier transformation along the wavenumber axis, an absolute optical path difference is obtained for each pixel independently of the other pixels in the field of view. As a result, interferograms with spatially distinct regions are analysed as easily as continuous ones. The approach is illustrated with a HSI system to measure 3-D profiles of optically smooth or rough surfaces. Compared to existing profilometers able to measure absolute path differences, the single shot nature of the approach provides greater immunity from environmental disturbance.

Computational hyperspectral interferometry for studies of brain function: proof of concept

Hyperspectral interferometric microscopy uses a unique combination of optics and algorithm design to extract information. Local brain activity rapidly changes local blood flow and red blood cell concentration (absorption) and oxygenation (color). We demonstrate that brain activity evoked during whisker stimulation can be detected with hyperspectral interferometric microscopy to identify the active whisker-barrel cortex in the rat brain. Information about constituent components is extracted across the entire spectral band. Algorithms can be flexibly optimized to discover, detect, quantify, and visualize a wide range of significant biological events, including changes relevant to the diagnosis and treatment of disease.