Spectral characterization of near-infrared acousto-optic tunable filter (AOTF) hyperspectral imaging systems using standard calibration materials

In this study, we propose and evaluate a method for spectral characterization of acousto-optic tunable filter (AOTF) hyperspectral imaging systems in the near-infrared (NIR) spectral region from 900 nm to 1700 nm. The proposed spectral characterization method is based on the SRM-2035 standard reference material, exhibiting distinct spectral features, which enables robust non-rigid matching of the acquired and reference spectra. The matching is performed by simultaneously optimizing the parameters of the AOTF tuning curve, spectral resolution, baseline, and multiplicative effects. In this way, the tuning curve (frequency-wavelength characteristics) and the corresponding spectral resolution of the AOTF hyperspectral imaging system can be characterized simultaneously. Also, the method enables simple spectral characterization of the entire imaging plane of hyperspectral imaging systems. The results indicate that the method is accurate and efficient and can easily be integrated with systems operating in diffuse reflection or transmission modes. Therefore, the proposed method is suitable for characterization, calibration, or validation of AOTF hyperspectral imaging systems.     

Hyperspectral imager, from ultraviolet to visible, with a KDP acousto-optic tunable filter

Hyperspectral imaging in the ultraviolet to visible spectral region has applications in astronomy, biology, chemistry, medical sciences, etc. A novel electronically tunable, random-wavelength access, compact, no-moving-parts, vibration-insensitive, computer-controlled hyperspectral imager operating from 220 to 480 nm with a spectral resolution of 160 cm(-1), e.g., 2 nm at 350 nm, has been developed by use of a KDP acousto-optic tunable filter (AOTF) with an enhanced CCD camera and a pair of crossed calcite Glan-Taylor polarizing prisms. The linear and angular apertures of the AOTF are 1.5 x 1.5 cm2 and 1.2 degrees, respectively. Imager setup and spectral imaging results as well as analyses and discussion of various factors affecting image quality are presented.     

Visible reflectance hyperspectral imaging: characterization of a noninvasive,in vivo system for determining tissue perfusion

We characterize a visible reflectance hyperspectral imaging system for noninvasive, in vivo, quantitative analysis of human tissue in a clinical environment. The subject area is illuminated with a quartz-tungsten-halogen light source, and the reflected light is spectrally discriminated by a liquid crystal tunable filter (LCTF) and imaged onto a silicon charge-coupled device detector. The LCTF is continuously tunable within its useful visible spectral range (525-725 nm) with an average spectral full width at half-height bandwidth of 0.38 nm and an average transmittance of 10.0%. A standard resolution target placed 5.5 ft from the system results in a field of view with a 17-cm diameter and an optimal spatial resolution of 0.45 mm. The measured reflectance spectra are quantified in terms of apparent absorbance and formatted as a hyperspectral image cube. As a clinical example, we examine a model of vascular dysfunction involving both ischemia and reactive hyperemia during tissue reperfusion. In this model, spectral images, based upon oxyhemoglobin and deoxyhemoblobin signals in the 525-645-nm region, are deconvoluted using a multivariate least-squares regression analysis to visualize the spatial distribution of the percentages of oxyhemoglobin and deoxyhemoglobin in specific skin tissue areas.     

Liquid-crystal tunable filter spectral imaging for brain tumor demarcation

Past studies have demonstrated that combined fluorescence and diffuse reflectance spectroscopy can successfully discriminate between normal, tumor core, and tumor margin tissues in the brain. To achieve efficient, real-time surgical resection guidance with optical biopsy, probe-based spectroscopy must be extended to spectral imaging to spatially demarcate the tumor margins. We describe the design and characterization of a combined fluorescence and diffuse reflectance imaging system that uses liquid-crystal tunable filter technology. Experiments were conducted to quantitatively determine the linearity, field of view, spatial and spectral resolution, and wavelength sensitivity of the imaging system. Spectral images were acquired from tissue phantoms, mouse brain in vitro, and human cortex in vivo for functional testing of the system. The spectral imaging system produces measured intensities that are linear with sample emission intensity and integration time and possesses a 1 in. (2.54 cm) field of view for a 7 in. (18 cm) object distance. The spectral resolution is linear with wavelength, and the spatial resolution is pixel-limited. The sensitivity spectra for the imaging system provide a guide for the distribution of total image integration time between wavelengths. Functional tests in vitro demonstrate the capability to spectrally discriminate between brain tissues based on exogenous fluorescence contrast or endogenous tissue composition. In vivo imaging captures adequate fluorescence and diffuse reflectance intensities within a clinically viable 2 min imaging time frame and demonstrates the importance of hemostasis to acquired signal strengths and imaging speed.     

Finite conjugate embedded relay lens hyperspectral imaging system (ERL-HIS)

We present a novel embedded relay lens hyperspectral imaging system (ERL-HIS) with high spectral resolution (nominal spectral resolution of 2.8 nm) and spatial resolution (30 μm×8 μm) that transfers the scanning plane to an additional imaging plane through the internal relay lens so as to alleviate all outside moving parts for the scanning mechanism used in the traditional HIS, where image scanning is achieved by the relative movement between the object and hyperspectrometer. The ERL-HIS also enables high-speed scanning and can attach to a variety of optical modules for versatile applications. Here, we also demonstrate an application of the proposed ERL-HIS attached to a microscopic system for observing autofluorescent images of sliced cancer tissue samples.     

Coherent array imaging using phased subarrays. Part II: simulations and experimental results

The basic principles and theory of phased subarray (PSA) imaging imaging provides the flexibility of reducing the number of front-end hardware channels between that of classical synthetic aperture (CSA) imaging--which uses only one element per firing event--and full-phased array (FPA) imaging-which uses all elements for each firing. The performance of PSA generally ranges between that obtained by CSA and FPA using the same array, and depends on the amount of hardware complexity reduction. For the work described in this paper, we performed FPA, CSA, and PSA imaging of a resolution phantom using both simulated and experimental data from a 3-MHz, 3.2-cm, 128-element capacitive micromachined ultrasound transducer (CMUT) array. The simulated system point responses in the spatial and frequency domains are presented as a means of studying the effects of signal bandwidth, reconstruction filter size, and subsampling rate on the PSA system performance. The PSA and FPA sector-scanned images were reconstructed using the wideband experimental data with 80% fractional bandwidth, with seven 32-element subarrays used for PSA imaging. The measurements on the experimental sector images indicate that, at the transmit focal zone, the PSA method provides a 10% improvement in the 6-dB lateral resolution, and the axial point resolution of PSA imaging is identical to that of FPA imaging. The signal-to-noise ratio (SNR) of PSA image was 58.3 dB, 4.9 dB below that of the FPA image, and the contrast-to-noise ratio (CNR) is reduced by 10%. The simulated and experimental test results presented in this paper validate theoretical expectations and illustrate the flexibility of PSA imaging as a way to exchange SNR and frame rate for simplified front-end hardware.     

Raman microspectroscopy: a comparison of point, line, and wide-field imaging methodologies

Three different Raman microspectroscopic imaging methodologies using a single experimental configuration are compared; namely, point and line mapping, as representatives of serial imaging approaches, and direct or wide-field Raman imaging employing liquid-crystalline tunable filters are surveyed. Raman imaging data acquired with equivalent low-power 514.5-nm laser excitation and a cooled CCD camera are analyzed with respect to acquisition times, image quality, spatial resolution, intensity profiles along spatial coordinates, and spectral signal-to-noise ratios (SNRs). Point and line mapping techniques provide similar SNRs and reconstructed Raman images at spatial resolutions of approximately 1.1 microm. In contrast, higher spatial resolution is obtained by direct, global imaging (approximately 313 nm), allowing subtle morphological features on test samples to be resolved.     

Instrumental error in chromotomosynthetic hyperspectral imaging

Chromotomosynthetic imaging (CTI) is a method of convolving spatial and spectral information that can be reconstructed into a hyperspectral image cube using the same transforms employed in medical tomosynthesis. A direct vision prism instrument operating in the visible (400-725?nm) with 0.6?mrad instantaneous field of view (IFOV) and 0.6-10?nm spectral resolution has been constructed and characterized. Reconstruction of hyperspectral data cubes requires an estimation of the instrument component properties that define the forward transform. We analyze the systematic instrumental error in collected projection data resulting from prism spectral dispersion, prism alignment, detector array position, and prism rotation angle. The shifting and broadening of both the spectral lineshape function and the spatial point spread function in the reconstructed hyperspectral imagery is compared with experimental results for monochromatic point sources. The shorter wavelength (λ<500 nm) region where the prism has the highest spectral dispersion suffers mostly from degradation of spectral resolution in the presence of systematic error, while longer wavelengths (λ>600 nm) suffer mostly from a shift of the spectral peaks. The quality of the reconstructed hyperspectral imagery is most sensitive to the misalignment of the prism rotation mount. With less than 1° total angular error in the two axes of freedom, spectral resolution was degraded by as much as a factor of 2 in the blue spectral region. For larger errors than this, spectral peaks begin to split into bimodal distributions, and spatial point response functions are reconstructed in rings with radii proportional to wavelength and spatial resolution.     

高光谱成像与应用技术发展

介绍了高光谱成像的需求与应用、光谱成像技术的现状与发展。高光谱成像已在产品分选、精准农业、环境监测、文物保护、刑事侦查、伪装识别等行业得到应用。传统的棱镜光栅色散型、连续可调谐滤光型、傅里叶变换干涉型等光谱成像分光方式成本高、体积大、速度慢,目前其光谱成像仪仍作为主要的研究设备。为促进产业化应用,需要发展体积小、成本低、速度快的光谱成像技术。计算层析、压缩编码、胶体量子点CQDs光谱成像技术仍需理论突破,短时间难以实用。积分视场、离散采样光谱成像技术原理简单、技术成熟,可用于空间分辨率要求不高的场合。复眼滤光式、像素滤光式光谱成像技术通过单片集成像素级滤光片,既能在权衡光谱分辨率与空间分辨率的基础上实现实时性,又能极大地减小体积、降低成本。     

Hyperspectral confocal microscope

We have developed a new, high performance, hyperspectral microscope for biological and other applications. For each voxel within a three-dimensional specimen, the microscope simultaneously records the emission spectrum from 500 nm to 800 nm, with better than 3 nm spectral resolution. The microscope features a fully confocal design to ensure high spatial resolution and high quality optical sectioning. Optical throughput and detection efficiency are maximized through the use of a custom prism spectrometer and a backside thinned electron multiplying charge coupled device (EMCCD) array. A custom readout mode and synchronization scheme enable 512-point spectra to be recorded at a rate of 8300 spectra per second. In addition, the EMCCD readout mode eliminates curvature and keystone artifacts that often plague spectral imaging systems. The architecture of the new microscope is described in detail, and hyperspectral images from several specimens are presented.     

Phase grating design for a dual-band snapshot imaging spectrometer

Infrared spectral features have proved useful in the identification of threat objects. Dual-band focal-plane arrays (FPAs) have been developed in which each pixel consists of superimposed midwave and long-wave photodetectors [Dyer and Tidrow, Conference on Infrared Detectors and Focal Plane Arrays (SPIE, Bellingham, Wash., 1999), pp. 434-440]. Combining dual-band FPAs with imaging spectrometers capable of interband hyperspectral resolution greatly improves spatial target discrimination. The computed-tomography imaging spectrometer (CTIS) [Descour and Dereniak, Appl. Opt. 34, 4817-4826 (1995)] has proved effective in producing hyperspectral images in a single spectral region. Coupling the CTIS with a dual-band detector can produce two hyperspectral data cubes simultaneously. We describe the design of two-dimensional, surface-relief, computer-generated hologram dispersers that permit image information in these two bands simultaneously.     

Experimental results from an airborne static Fourier transform imaging spectrometer

A high étendue static Fourier transform spectral imager has been developed for airborne use. This imaging spectrometer, based on a Michelson interferometer with rooftop mirrors, is compact and robust and benefits from a high collection efficiency. Experimental airborne images were acquired in the visible domain. The processing chain to convert raw images to hyperspectral data is described, and airborne spectral images are presented. These experimental results show that the spectral resolution is close to the one expected, but also that the signal to noise ratio is limited by various phenomena (jitter, elevation fluctuations, and one parasitic image). We discuss the origin of those limitations and suggest solutions to circumvent them.     

Implementation of a 3-D Hyperspectral Instrument for Skin Imaging Applications

The many bands acquired by typical multispectral, hyperspectral, and ultraspectral instruments are collected in either a scanning or staring fashion. Staring instruments, like that described in this paper, are popular because they are capable of producing spatially coherent images of a target scene; therefore, they are suitable for noncontact inspection applications. However, the wavebands need to be coregistered. A new application of hyperspectral instrumentation is proposed, where a sheet-of-light method, produced by a low-power laser light, is used to compute range measurements, and a hyperspectral instrument is used to acquire spectral information in the visible (VIS) range of the spectrum. The main advantage of this method is that a single hyperspectral tunable filter arrangement is used to sweep the image to get 3-D information and acquire hyperspectral spectral measurements of a scene. This paper describes the implementation of the image acquisition subsystem, which was done in LabView, and the method used to obtain coregistered radiometrically calibrated spectral imagery and range information. Test cases showed that the instrument may be used to retrieve height and spectral information of small areas and could be used for skin-imaging applications.      

Spatial-spectral modulating snapshot hyperspectral imager

Experimental results are presented for a computed tomography imaging spectrometer (CTIS) with imposed spatial-spectral modulation on the image scene. This modulation structure on the CTIS tomographic dispersion created substantial gains in spectral reconstruction resolution after standard iterative, nonlinear, inversion techniques were used. Modulation limits system ambiguities, so high-frequency spectral and low-frequency spatial scene data could be recovered. The results demonstrate how spatial modulation acts as a high-frequency spectral deconvolver for the snapshot hyperspectral imager technology.     

Active DLP hyperspectral illumination: a noninvasive, in vivo, system characterization visualizing tissue oxygenation at near video rates

We report use of a novel hyperspectral imaging system utilizing digital light processing (DLP) technology to noninvasively visualize in vivo tissue oxygenation during surgical procedures. The system's novelty resides in its method of illuminating tissue with precisely predetermined continuous complex spectra. The Texas Instruments digital micromirror device, DMD, chip consisting of 768 by 1024 mirrors, each 16 μm square, can be switched between two positions at 12.5 kHz. Switching the appropriate mirrors controls the intensity of light illuminating the tissue as a function of wavelength, active spectral illumination. Meaning, the tissue can be illuminated with a different spectrum of light within 80 μs. Precisely, predetermined spectral illumination penetrates into patient tissue, its chemical composition augments the spectral properties of the light, and its reflected spectra are detected and digitized at each pixel detector of a silicon charge-coupled device, CCD. Using complex spectral illumination, digital signal processing and chemometric methods produce chemically relevant images at near video rates. Specific to this work, tissue is illuminated spectrally with light spanning the visible electromagnetic spectrum (380 to 780 nm). Spectrophotometric images are detected and processed visualizing the percentage of oxyhemoglobin at each pixel detector and presented continuously, in real time, at 3 images per second. As a proof of principle application, kidneys of four live anesthetized pigs were imaged before, during, and after renal vascular occlusion. DLP Hyperspectral Imaging with active spectral illumination detected a 64.73 ± 1.5% drop in the oxygenation of hemoglobin within 30 s of renal arterial occlusion. Producing chemically encoded images at near video rate, time-resolved hyperspectral imaging facilitates monitoring renal blood flow during animal surgery and holds considerable promise for doing the same during human surgical interventions.     

Analysis of hyperspectral fluorescence images for poultry skin tumor inspection

We present a hyperspectral fluorescence imaging system with a fuzzy inference scheme for detecting skin tumors on poultry carcasses. Hyperspectral images reveal spatial and spectral information useful for finding pathological lesions or contaminants on agricultural products. Skin tumors are not obvious because the visual signature appears as a shape distortion rather than a discoloration. Fluorescence imaging allows the visualization of poultry skin tumors more easily than reflectance. The hyperspectral image samples obtained for this poultry tumor inspection contain 65 spectral bands of fluorescence in the visible region of the spectrum at wavelengths ranging from 425 to 711 nm. The large amount of hyperspectral image data is compressed by use of a discrete wavelet transform in the spatial domain. Principal-component analysis provides an effective compressed representation of the spectral signal of each pixel in the spectral domain. A small number of significant features are extracted from two major spectral peaks of relative fluorescence intensity that have been identified as meaningful spectral bands for detecting tumors. A fuzzy inference scheme that uses a small number of fuzzy rules and Gaussian membership functions successfully detects skin tumors on poultry carcasses. Spatial-filtering techniques are used to significantly reduce false positives.     

Some practical issues in anomaly detection and exploitation of regions of interest in hyperspectral images

We address method of detection of anomalies in hyperspectral images that consists in performing the detection when the spectral signatures of the targets are unknown. We show that, in real hyperspectral images, use of the full spectral resolution may not be necessary for detection but that the correlation properties of spectral fluctuations have to be taken into account in the design of the detection algorithm. Anomaly detectors are useful for detecting regions of interest (ROIs), but, as they are prone to false alarms, one must analyze the ROIs obtained further to decide whether they correspond to real targets. We propose a method of exploitation of these ROIs that consists in generating a single image in which the contrast of the ROI is optimized.     

Time-resolved diffuse optical tomographic imaging for the provision of both anatomical and functional information about biological tissue

We present in vivo images of near-infrared (NIR) diffuse optical tomography (DOT) of human lower legs and forearm to validate the dual functions of a time-resolved (TR) NIR DOT in clinical diagnosis, i.e., to provide anatomical and functional information simultaneously. The NIR DOT system is composed of time-correlated single-photon-counting channels, and the image reconstruction algorithm is based on the modified generalized pulsed spectral technique, which effectively incorporates the TR data with reasonable computation time. The reconstructed scattering images of both the lower legs and the forearm revealed their anatomies, in which the bones were clearly distinguished from the muscles. In the absorption images, some of the blood vessels were observable. In the functional imaging, a subject was requested to do handgripping exercise to stimulate physiological changes in the forearm tissue. The images of oxyhemoglobin, deoxyhemoglobin, and total hemoglobin concentration changes in the forearm were obtained from the differential images of the absorption at three wavelengths between the exercise and the rest states, which were reconstructed with a differential imaging scheme. These images showed increases in both blood volume and oxyhemoglobin concentration in the arteries and simultaneously showed hypoxia in the corresponding muscles. All the results have demonstrated the capability of TR NIR DOT by reconstruction of the absolute images of the scattering and the absorption with a high spatial resolution that finally provided both the anatomical and functional information inside bulky biological tissues.     

基于显微高光谱成像的人血细胞研究

使用自行研制的推帚式显微高光谱成像系统采集了正常,白血病人血液涂片的显微高光谱图像数据.通过对正常、白血病人血液显微高光谱数据进行处理,获得了人血液的单波段图像,并提取了部分血液细胞的典型透射率光谱曲线.分析这些曲线发现,病变细胞透射率普遍高于正常细胞,特别是在541.3 nm附近透射率高出了50%左右.通过对血液涂片的图像和光谱特征进行分析表明,经过一定的改进,可以将显微高光谱成像系统作为一种新的检测手段,辅助医学研究人员对人血液进行分析.     

III-Nitride Heterostructure Layered Tunnel Barriers For a Tunable Hyperspectral Detector

We report on the fabrication and characterization of Ill-nitride layered tunnel barriers with applications for a new type of tunable hyperspectral imaging detector with intrinsically hyperspectral pixels. This would enable each pixel to be individually tunable in real-time through a range of wavelengths, with the number and width of spectral channels being dynamically adjustable. Shape-engineered electron barriers fabricated from III-nitride heterostructures allow barrier height to be varied by application of a voltage. A spectroscopy of photon wavelength is enabled via the collection of photoexcited electrons across this barrier. The device is envisioned for tunable detection of ultraviolet through infrared wavelengths.