Image Acquisition Speed Research Articles

High-Throughput Cell Imaging and Classification by Narrowband and Low-Spectral-Resolution Raman Microscopy.

We investigated the use of narrowband Raman spectra for rapid label-free molecular imaging aimed at cell classification using principal component regression and linear discriminant analysis. In the classification of breast nontumorigenic epithelial and cancer cell lines, the classification accuracies using a spectral range of 100 cm-1 were equivalent to or better than that with using the fingerprint and high-wavenumber regions. Narrowing the Raman spectral range for analysis allows reduction of the charge-coupled device (CCD) pixels required for spectrum detection, resulting in the improvement of image acquisition speed with adequate classification accuracy. Our measurements revealed that the wavenumber region at 1397-1501 cm-1 can provide molecular information sufficient for cell classification without causing notable errors in the baseline-correction. A spectral resolution of ∼9 cm-1 was found to be sufficient to provide high accuracy in cell classification, which allowed us to apply pixel binning at the CCD readout for further acceleration of the imaging speed. As a result, the acquisition time for a 1200 × 1500 pixels Raman hyperspectral image at 1397-1501 cm-1 was reduced to 21 min. Under this condition, different cell lines were classified at accuracies higher than 90%. The presented approach will improve throughput of cell and tissue analysis and classification using Raman spectroscopy and extend practical uses of Raman imaging in biology and medicine.

Development of an artificial cloud lighting condition system using machine vision for strawberry powdery mildew disease detection

Strawberry plants have been facing a significant proportion of diseases during cultivation, scattered throughout the field, emphasizing the need for proper diseases management. Powdery mildew is one of the major fungal strawberry disease which is typically responsible for approximately 30–70% loss of yields. The aim of this study was to develop a machine vision based artificial cloud lighting condition system for detecting strawberry powdery mildew leaf disease. The artificial cloud lighting condition system was developed consisting of custom software, two µEye colour cameras, a black cloth cover, real time kinematics-global positioning system and a ruggedized laptop computer and mounted on a mobile platform. The custom software was developed in C# programming language. The colour co-occurrence matrix based texture analysis was used to extract image features and discriminant analysis (quadratic) for classification. The study proposed mobile platform of artificial cloud lighting condition for image acquisition is beneficial. It showed higher detection accuracies of 95.26%, 95.45% and 95.37% for recall, precision and F-measure, respectively compared to 81.54%, 72% and 75.95% of recall, precision and F-measure, respectively with acquired images at natural cloud lighting condition. The feature selection results suggested the PM_GHSI feature model was best fit for this study. This study also revealed that the image acquisition speed (1.5 km h−1) and working depth (300 mm) are suitable for strawberry powdery mildew disease detection in real-time field condition.

TCR microclusters form spatially segregated domains and sequentially assemble in calcium-dependent kinetic steps

Engagement of the T cell receptor (TCR) by stimulatory ligand results in the rapid formation of microclusters at sites of T cell activation. Whereas microclusters have been studied extensively using confocal microscopy, the spatial and kinetic relationships of their signaling components have not been well characterized due to limits in image resolution and acquisition speed. Here we show, using TIRF-SIM to examine the organization of microclusters at sub-diffraction resolution, the presence of two spatially distinct domains composed of ZAP70-bound TCR and LAT-associated signaling complex. Kinetic analysis of microcluster assembly reveal surprising delays between the stepwise recruitment of ZAP70 and signaling proteins to the TCR, as well as distinct patterns in their disassociation. These delays are regulated by intracellular calcium flux downstream of T cell activation. Our results reveal novel insights into the spatial and kinetic regulation of TCR microcluster formation and T cell activation.

Deep Group-Wise Registration for Multi-Spectral Images From Fundus Images

Multi-spectral imaging (MSI) is a novel non-invasive tool for visualizing the entire span of the eye, from the internal limiting membrane to the choroid. However, spatial misalignments can be frequently observed in sequential MSI images because the eye saccade movement is usually faster than the MSI image acquisition speed. Therefore, registering MSI images is necessary for computer-based analysis of retinal degeneration via MSI. In this paper, we propose an early deep learning framework for achieving an accurate registration of MSI images in a group-wise fashion. The framework contains three parts: a template construction based on principal component analysis, a deformation field calculation, and a spatial transformation. The framework is uniquely capable of resolving two key challenges, i.e., the “multi-modal” characteristics in MSI images for the acquisition with different spectra and the requirement of joint registration of the sequential images. Our experimental results demonstrate the superior performance of our framework compared to several representative state-of-the-art techniques in both speed and accuracy.

Using elongated microparticles to enhance tailorable nanoemulsion delivery in excised human skin and volunteers

This study demonstrates, for the first time, clinical testing of elongated silica microparticles (EMP) combined with tailorable nanoemulsions (TNE) to enhance topical delivery of hydrophobic drug surrogates. Likewise, this is the first report of 6-carboxyfluorescein (a model molecule for topically delivered hydrophobic drugs) AM1 & DAMP4 (novel short peptide surfactants) used in volunteers. The EMP penetrates through the epidermis and stop at the dermal-epidermal junction (DEJ). TNE are unusually stable and useful because the oil core allows high drug loading levels and the surface properties can be easily controlled. At first, we chose alginate as a crosslinking agent between EMP and TNE. We initially incorporated a fluorescent lipophilic dye, DiI, as a hydrophobic drug surrogate into TNE for visualization with microscopy. We compared four different coating approaches to combine EMP and TNE and tested these formulations in freshly excised human skin. The delivery profile characterisation was imaged by dye- free coherent anti-Stoke Raman scattering (CARS) microscopy to detect the core droplet of TNE that was packed with pharmaceutical grade lipid (glycerol) instead of DiI. These data show the EMP penetrating to the DEJ followed by controlled release of the TNE. Freeze-dried formulations with crosslinking resulted in a sustained release profile, whereas a freeze-dried formulation without crosslinking showed an immediate burst-type release profile. Finally, we tested the crosslinked TNE coated EMP formulation in volunteers using multiphoton microscopy (MPM) and fluorescence-lifetime imaging microscopy (FLIM) to document the penetration depth characteristics. These forms of microscopy have limitations in terms of image acquisition speed and imaging area coverage but can detect fluorescent drug delivery through the superficial skin in volunteers. 6-Carboxyfluorescein was selected as the fluorescent drug surrogate for the volunteer study based on the similarity of size, charge and hydrophobicity characteristics to small therapeutic drugs that are difficult to deliver through skin. The imaging data showed a 6-carboxyfluorescein signal deep in volunteer skin supporting the hypothesis that EMP can indeed enhance the delivery of TNE in human skin. There were no adverse events recorded at the time of the study or after the study, supporting the use of 6-carboxyfluorescein as a safe and detectable drug surrogate for topical drug research. In conclusion, dry formulations, with controllable release profiles can be obtained with TNE coated EMP that can effectively enhance hydrophobic payload delivery deep into the human epidermis.

Rapid 3D image scanning microscopy with multi-spot excitation and double-helix point spread function detection.

Refocusing after Scanning using Helical phase engineering (RESCH) microscopy has previously been demonstrated to provide volumetric information from a single 2D scan. However, the practical application of this method is challenging due to its limited image acquisition speed and spatial resolution. Here, we report on a combination of RESCH and multifocal structured illumination microscopy (MSIM) to improve the image acquisition speed and spatial resolution. A phase mask is introduced to modulate the conventional point spread function (PSF) to the double-helix PSF (DH-PSF), which provides volumetric information, and meanwhile, sparse multifocal illumination patterns are generated by a digital micromirror device (DMD) for parallel 3D subdiffractive imaging information acquisition. We also present a strategy for processing the collected raw data with a Richardson-Lucy deconvolution and pixel reassignment algorithm to improve the spatial resolution of the depth estimation and imaging performance. The proposed 3D image scanning microscopy can record 3D specimen information and the corresponding depth information from a single multi-spot 2D planar scan, which ensures faster data acquisition, larger field of view, and higher spatial resolution than RESCH. Finally, we demonstrate the capability of our system with actual experiments.

High speed CMOS acquisition system based on FPGA embedded image processing for electro-optical measurements

Electro-optical measurements, i.e., optical waveguides and plasmonic based electrochemical impedance spectroscopy (P-EIS), are based on the sensitive dependence of refractive index of electro-optical sensors on surface charge density, modulated by an AC electrical field applied to the sensor surface. Recently, P-EIS has emerged as a new analytical tool that can resolve local impedance with high, optical spatial resolution, without using microelectrodes. This study describes a high speed image acquisition and processing system for electro-optical measurements, based on a high speed complementary metal-oxide semiconductor (CMOS) sensor and a field-programmable gate array (FPGA) board. The FPGA is used to configure CMOS parameters, as well as to receive and locally process the acquired images by performing Fourier analysis for each pixel, deriving the real and imaginary parts of the Fourier coefficients for the AC field frequencies. An AC field generator, for single or multi-sine signals, is synchronized with the high speed acquisition system for phase measurements. The system was successfully used for real-time angle-resolved electro-plasmonic measurements from 30 Hz up to 10 kHz, providing results consistent to ones obtained by a conventional electrical impedance approach. The system was able to detect amplitude variations with a relative variation of ±1%, even for rather low sampling rates per period (i.e., 8 samples per period). The PC (personal computer) acquisition and control software allows synchronized acquisition for multiple FPGA boards, making it also suitable for simultaneous angle-resolved P-EIS imaging.

Quality assessment of the optical thin films using line field spectral domain optical coherence tomography

In this study, the industrial inspection of optical thin film on touch screen panels was demonstrated using line field spectral domain optical coherence tomography. The conventional Fourier domain optical coherence tomography system requires a single scanner for two-dimensional cross-sectional images and two scanners for volumetric images. Our developed line field spectral domain optical coherence tomography has the advantage of needing only a single scanner for volumetric images, while two-dimensional cross-sectional images are obtained by the parallel acquisition of an illuminated line on a sample using an area camera. Further, the image acquisition speed was enhanced by implementing a high speed camera (340 frames per seconds) with improved quantum efficiency at near infrared region enabling two-fold frame rate. Cross-sectional and volumetric images were acquired to detect the internal sublayer defects in the optical thin films, which are difficult to observe using visual or machine vision-based inspection methods. The developed pseudo code for defect identification in optical thin films was well-utilized here for the defect inspection. The system characterization is demonstrated using United State Air Force (USAF) resolution target. The results indicate the possible application of the proposed system in touch screen panel inspection for the quality assurance of products at the consumer end.

Pool boiling of ethanol and FC-72 on open microchannel surfaces

The paper presents experimental investigations into pool boiling heat transfer for open microchannel surfaces. Parallel microchannels fabricated by machining were about 0.3 mm wide, and 0.2 to 0.5 mm deep and spaced every 0.1 mm. The experiments were carried out for ethanol, and FC-72 at atmospheric pressure. The image acquisition speed was 493 fps (at resolution 400 × 300 pixels with Photonfocus PHOT MV-D1024-160-CL camera). Visualization investigations aimed to identify nucleation sites and flow patterns and to determine the bubble departure diameter and frequency at various superheats. The primary factor in the increase of heat transfer coefficient at increasing heat flux was a growing number of active pores and increased departure frequency. Heat transfer coefficients obtained in this study were noticeably higher than those from a smooth surface.

A real-time and registration-free framework for dynamic shape instantiation.

Real-time 3D navigation during minimally invasive procedures is an essential yet challenging task, especially when considerable tissue motion is involved. To balance image acquisition speed and resolution, only 2D images or low-resolution 3D volumes can be used clinically. In this paper, a real-time and registration-free framework for dynamic shape instantiation, generalizable to multiple anatomical applications, is proposed to instantiate high-resolution 3D shapes of an organ from a single 2D image intra-operatively. Firstly, an approximate optimal scan plane was determined by analyzing the pre-operative 3D statistical shape model (SSM) of the anatomy with sparse principal component analysis (SPCA) and considering practical constraints. Secondly, kernel partial least squares regression (KPLSR) was used to learn the relationship between the pre-operative 3D SSM and a synchronized 2D SSM constructed from 2D images obtained at the approximate optimal scan plane. Finally, the derived relationship was applied to the new intra-operative 2D image obtained at the same scan plane to predict the high-resolution 3D shape intra-operatively. A major feature of the proposed framework is that no extra registration between the pre-operative 3D SSM and the synchronized 2D SSM is required. Detailed validation was performed on studies including the liver and right ventricle (RV) of the heart. The derived results (mean accuracy of 2.19mm on patients and computation speed of 1ms) demonstrate its potential clinical value for real-time, high-resolution, dynamic and 3D interventional guidance.

A Fully Integrated Dual-Channel On-Coil CMOS Receiver for Array Coils in 1.5-10.5 T MRI.

Magnetic resonance imaging (MRI) is among the most important medical imaging modalities. Coil arrays and receivers with high channel counts (16 and more) have to be deployed to obtain the image quality and acquisition speed required by modern clinical protocols. In this paper, we report the theoretical analysis, the system-level design, and the circuit implementation of the first receiver IC (RXIC) for clinical MRI fully integrated in a modern CMOS technology. The dual-channel RXIC sits directly on the sensor coil, thus eliminating any RF cable otherwise required to transport the information out of the magnetic field. The first stage LNA was implemented using a noise-canceling architecture providing a highly reflective input used to decouple the individual channels of the array. Digitization is performed directly on-chip at base-band by means of a delta-sigma modulator, allowing the subsequent optical transmission of data. The presented receiver, implemented in a CMOS technology, is compatible with MRI scanners up to . It reaches sub- noise figure for MRI units and features a dynamic range up to at a power consumption below per channel, with an area occupation of . Mounted on a small-sized printed circuit board (PCB), the receiver IC has been employed in a commercial MRI scanner to acquire in-vivo images matching the quality of traditional systems, demonstrating the first step toward multichannel wearable MRI array coils.

Characterization of an industry-grade CMOS camera well suited for single molecule localization microscopy \u2013 high performance super-resolution at low cost

Many commercial as well as custom-built fluorescence microscopes use scientific-grade cameras that represent a substantial share of the instrument’s cost. This holds particularly true for super-resolution localization microscopy where high demands are placed especially on the detector with respect to sensitivity, noise, and also image acquisition speed. Here, we present and carefully characterize an industry-grade CMOS camera as a cost-efficient alternative to commonly used scientific cameras. Direct experimental comparison of these two detector types shows widely similar performance for imaging by single molecule localization microscopy (SMLM). Furthermore, high image acquisition speeds are demonstrated for the CMOS detector by ultra-fast SMLM imaging.

Fast Fourier single-pixel imaging via binary illumination

Fourier single-pixel imaging (FSI) employs Fourier basis patterns for encoding spatial information and is capable of reconstructing high-quality two-dimensional and three-dimensional images. Fourier-domain sparsity in natural scenes allows FSI to recover sharp images from undersampled data. The original FSI demonstration, however, requires grayscale Fourier basis patterns for illumination. This requirement imposes a limitation on the imaging speed as digital micro-mirror devices (DMDs) generate grayscale patterns at a low refreshing rate. In this paper, we report a new strategy to increase the speed of FSI by two orders of magnitude. In this strategy, we binarize the Fourier basis patterns based on upsampling and error diffusion dithering. We demonstrate a 20,000 Hz projection rate using a DMD and capture 256-by-256-pixel dynamic scenes at a speed of 10 frames per second. The reported technique substantially accelerates image acquisition speed of FSI. It may find broad imaging applications at wavebands that are not accessible using conventional two-dimensional image sensors.

Fast two-snapshot structured illumination for temporal focusing microscopy with enhanced axial resolution

We present a new two-snapshot structured light illumination (SLI) reconstruction algorithm for fast image acquisition. The new algorithm, which only requires two mutually π phase-shifted raw structured images, is implemented on a custom-built temporal focusing fluorescence microscope (TFFM) to enhance its axial resolution via a digital micromirror device (DMD). First, the orientation of the modulated sinusoidal fringe patterns is automatically identified via spatial frequency vector detection. Subsequently, the modulated in-focal-plane images are obtained via rotation and subtraction. Lastly, a parallel amplitude demodulation method, derived based on Hilbert transform, is applied to complete the decoding processes. To demonstrate the new SLI algorithm, a TFFM is custom-constructed, where a DMD replaces the generic blazed grating in the system and simultaneously functions as a diffraction grating and a programmable binary mask, generating arbitrary fringe patterns. The experimental results show promising depth-discrimination capability with an axial resolution enhancement factor of 1.25, which matches well with the theoretical estimation, i.e, 1.27. Imaging experiments on pollen grain and mouse kidney samples have been performed. The results indicate that the two-snapshot algorithm presents comparable contrast reconstruction and optical cross-sectioning capability than those adopting the conventional root-mean-square (RMS) reconstruction method. The two-snapshot method can be readily applied to any sinusoidally modulated illumination systems to realize high-speed 3D imaging as less frames are required for each in-focal-plane image restoration, i.e., the image acquisition speed is improved by 2.5 times for any two-photon systems.

THz Imaging Using Uncooled Wideband Direct Detection Focal Plane Arrays

In millimeter and submillimeter-wave radiometric imaging systems, a persistent goal is the increase in the speed of acquisition of the image while maintaining a high sensitivity. Typically, the highest sensitivity is achieved by cryogenically cooling the detectors, specifically in astronomical applications. However, for the purpose of low-cost imaging applications, it is desirable to operate at room temperature. Without cryogenically cooling, the electronic noise introduced by the detectors becomes dominant, making the detectors less sensitive. Resorting to detection architectures containing amplification circuitry might be impractical for implementation in large focal plane arrays (FPAs) fabricated in integrated technologies. This contribution derives the focal plane architecture that maximizes the imaging speed of radiometers operating at room temperature without using any amplification circuitry. It is shown that in such scenario a practical image acquisition speed can still be achieved when a very broad portion of the THz-band is exploited. Ultimately, the imaging speed is maximized when the FPA is undersampled, implying a tradeoff in the size of the optics. The analysis is substantiated by a case study with recently developed wideband leaky lens antenna feeds operating from 200 to 600 GHz.

In situ high-resolution thermal microscopy on integrated circuits

The miniaturization of metal tracks in integrated circuits (ICs) can cause abnormal heat dissipation, resulting in electrostatic discharge, overvoltage breakdown, and other unwanted issues. Unfortunately, locating areas of abnormal heat dissipation is limited either by the spatial resolution or imaging acquisition speed of current thermal analytical techniques. A rapid, non-contact approach to the thermal imaging of ICs with sub-μm resolution could help to alleviate this issue. In this work, based on the intensity of the temperature-dependent two-photon fluorescence (TPF) of Rhodamine 6G (R6G) material, we developed a novel fast and non-invasive thermal microscopy with a sub-μm resolution. Its application to the location of hotspots that may evolve into thermally induced defects in ICs was also demonstrated. To the best of our knowledge, this is the first study to present high-resolution 2D thermal microscopic images of ICs, showing the generation, propagation, and distribution of heat during its operation. According to the demonstrated results, this scheme has considerable potential for future in situ hotspot analysis during the optimization stage of IC development.

Rotating single-shot acquisition (RoSA) with composite reconstruction for fast high-resolution diffusion imaging.

To accelerate high-resolution diffusion imaging, rotating single-shot acquisition (RoSA) with composite reconstruction is proposed. Acceleration was achieved by acquiring only one rotating single-shot blade per diffusion direction, and high-resolution diffusion-weighted (DW) images were reconstructed by using similarities of neighboring DW images. A parallel imaging technique was implemented in RoSA to further improve the image quality and acquisition speed. RoSA performance was evaluated by simulation and human experiments. A brain tensor phantom was developed to determine an optimal blade size and rotation angle by considering similarity in DW images, off-resonance effects, and k-space coverage. With the optimal parameters, RoSA MR pulse sequence and reconstruction algorithm were developed to acquire human brain data. For comparison, multishot echo planar imaging (EPI) and conventional single-shot EPI sequences were performed with matched scan time, resolution, field of view, and diffusion directions. The simulation indicated an optimal blade size of 48 × 256 and a 30 ° rotation angle. For 1 × 1 mm2 in-plane resolution, RoSA was 12 times faster than the multishot acquisition with comparable image quality. With the same acquisition time as SS-EPI, RoSA provided superior image quality and minimum geometric distortion. RoSA offers fast, high-quality, high-resolution diffusion images. The composite image reconstruction is model-free and compatible with various diffusion computation approaches including parametric and nonparametric analyses. Magn Reson Med 79:264-275, 2018. © 2017 International Society for Magnetic Resonance in Medicine.

Optimization and applications of an excitation-scanning hyperspectral imaging system.

Currently, the majority of microscopic and endoscopic technologies utilize white light illumination. For a number of applications, hyper-spectral imaging can be shown to have significant improvements over standard white-light imaging techniques. This is true for both microscopy and in vivo imaging. However, hyperspectral imaging methods have suffered from slow application times. Often, minutes are required to gather a full imaging stack. Here we will describe the system and evaluate optimizations and applications of a novel excitation-scanning hyperspectral imaging system. We have developed and are optimizing a novel approach called excitation-scanning hyperspectral imaging that provides an order of magnitude increased signal strength. Optimization of the light path, optical components and illumination sources have allowed us to achieve high speed image acquisition. This high speed allows for potential live video acquisition. This excitation-scanning hyperspectral imaging technology has potential to impact a range of applications. The current system allows triggering of up to 16 wavelengths at less than 1 millisecond per image using digital strobing. Analog intensity control is also provided for a fully customizable excitation profile. A significant advantage of excitation-scanning hyperspectral imaging is can identify multiple targets simultaneously in real time. We are optimizing the system to compare sensitivity and specificity of excitation-scanning hyperspectral imaging with pathology techniques. Finally, we are exploring utilizing this technology to measure cAMP distribution in three dimensions within a cell.

Fast demodulation of pattern images by spiral phase transform in structured-illumination reflectance imaging for detection of bruises in apples

A structured-illumination reflectance imaging (SIRI) system was recently developed in our laboratory for enhanced quality evaluation of horticultural products. It was implemented using a digital camera to acquire reflectance images from food products subjected to sinusoidal patterns of illumination, instead of conventional diffuse or uniform illumination. The reconstruction of amplitude images is a key step in implementing SIRI technique. Conventional methods require acquisition and subsequent demodulation of three phase-shifted pattern images, which limits the speed of image acquisition. This study proposed the use of spiral phase transform (SPT) for demodulation of pattern images acquired by SIRI. Three SPT-based methods involving only one or two images were investigated through numerical simulation, followed with experiments on the detection of fresh bruises in apples. Compared to conventional three-phase based and two-phase based Gram-Schmidt (GS) orthonormalization demodulation methods, the two-phase SPT method achieved the same or even higher demodulation accuracy, which applies to two pattern images that can be arbitrarily phase-shifted. SPT also allowed single-image based demodulation, although its performance still needs to be improved. This study demonstrated that SPT for amplitude demodulation can increase the imaging speed of SIRI by using one or two pattern images, which is a significant step towards real-time implementation of the technique.

MO-D-PinS Room/Hall E-01: Integrating MR into Radiation Therapy

MRI, with its excellent soft tissue contrast and its ability to provide physiological as well as anatomical information, is becoming increasingly used in radiation therapy for treatment planning, image-guided radiation therapy, and treatment evaluation. This session will explore solutions to integrating MRI into the simulation process. Obstacles for using MRI for simulation include distortions and artifacts, image acquisition speed, complexity of imaging techniques, and lack of electron density information. Partners in Solutions presents vendor representatives who will present their approaches to meeting these challenges and others. An increased awareness of how MRI simulation works will allow physicists to better understand and use this powerful technique. The speakers are all employees who are presenting information about their company’s products.