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Results for “multiplex imaging”

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At least 145 records · Page 8Linked to original sources

PubMed2002

Immunofluorescent labeling of cancer marker Her2 and other cellular targets with semiconductor quantum dots.

Semiconductor quantum dots (QDs) are among the most promising emerging fluorescent labels for cellular imaging. However, it is unclear whether QDs, which are nanoparticles rather than small molecules, can specifically and effectively label molecular targets at a subcellular level. Here we have used QDs linked to immunoglobulin G (IgG) and streptavidin to label the breast cancer marker Her2 on the surface of fixed and live cancer cells, to stain actin and microtubule fibers in the cytoplasm, and to detect nuclear antigens inside the nucleus. All labeling signals are specific for the intended targets and are brighter and considerably more photostable than comparable organic dyes. Using QDs with different emission spectra conjugated to IgG and streptavidin, we simultaneously detected two cellular targets with one excitation wavelength. The results indicate that QD-based probes can be very effective in cellular imaging and offer substantial advantages over organic dyes in multiplex target detection.

3T3 Cells↗

PubMed2006

Diffuse optical tomography system to image brain activation with improved spatial resolution and validation with functional magnetic resonance imaging.

Although most current diffuse optical brain imaging systems use only nearest- neighbor measurement geometry, the spatial resolution and quantitative accuracy of the imaging can be improved through the collection of overlapping sets of measurements. A continuous-wave diffuse optical imaging system that combines frequency encoding with time-division multiplexing to facilitate overlapping measurements of brain activation is described. Phantom measurements to confirm the expected improvement in spatial resolution and quantitative accuracy are presented. Experimental results showing the application of this instrument for imaging human brain activation are also presented. The observed improvement in spatial resolution is confirmed by functional magnetic resonance imaging.

Brain↗

PubMed2004

Real-time rectilinear 3-D ultrasound using receive mode multiplexing.

In previous work, we developed two generations of a real-time rectilinear volumetric scanner operating at 5 MHz for abdominal, breast, or vascular imaging using a Mills cross two-dimensional (2-D) array and a rectilinear periodic 2-D array. To improve spatial resolution performance and sensitivity, we developed a new design using 4:1 receive mode multiplexing. With 4:1 multiplexing, the new 65,000 element 2-D array has 4 x 256 = 1024 receivers so that 256 receivers can be used on any image line. The two major benefits of using receive mode multiplexing are an increase in receive sensitivity due to a greater number of receive elements, and a decrease in grating lobe and clutter levels due to increased receive element density. Theoretical simulations and analysis show an increase of about 13 dB in sensitivity compared to our previous work. With these encouraging results, a new 65,000 element 5-MHz, 2-D array having 1024 receivers and 169 transmitters was prototyped. In addition, the multiplexer and control circuitry were designed, built, and interfaced with both the transducer and volumetric scanner. Images of tissue-mimicking phantoms and in vivo targets were obtained. Using a spherical cyst phantom, experimental results showed a +12 dB improvement in signal-to-noise ratio and a +6 dB improvement in contrast compared to our previous work.

Adult↗

PubMed1985

Image reconstruction from coded data: I. Reconstruction algorithms and experimental results.

Two algorithms have been developed for reconstructing objects from their coded images and a priori knowledge of the object class. Reconstructions from both algorithms are presented, but the results appear to be largely independent of the algorithm used. One of the algorithms, a Monte Carlo approach, is used to investigate the quality of the reconstruction of two- and three-dimensional objects from simulated coded-image data with respect to viewing geometry and multiplexing (mixing) of the data. The cases examined include reconstructions from data with and without signal-dependent photon noise. It is found that reconstructing from multiplexed data is not so serious a problem as reconstructing from data obtained with a limited viewing angle. Also, when photon noise is included in the data, reconstructions obtained from multiplexed data are better than those obtained from unmultiplexed data because of the higher photon count made available by multiplexing. It appears that the fidelity of a reconstruction depends much more strongly on the design of the data-taking system (the coded apertures) than on the reconstruction algorithm.

Models, Structural↗

PubMed2005

Volume three-dimensional flow measurements using wavelength multiplexing.

Optically distinguishable seeding particles that emit light in a narrow bandwidth, and a combination of bandwidths, were prepared by encapsulating quantum dots. The three-dimensional components of the particles' displacement were measured within a volume of fluid with particle tracking velocimetry (PTV). Particles are multiplexed to different hue bands in the camera images, enabling an increased seeding density and (or) fewer cameras to be used, thereby increasing the measurement spatial resolution and (or) reducing optical access requirements. The technique is also applicable to two-phase flow measurements with PTV or particle image velocimetry, where each phase is uniquely seeded.

Algorithms↗

PubMed2005

Identification of Peptide tumor markers in a tumor graft model in immunodeficient mice.

The medical demand for useful biomarkers is large and still increasing. This is especially true for cancer, because for this disease adequate diagnostic markers with high specificity and sensitivity are still lacking. Despite advances in imaging technologies for early detection of cancer, peptidomic multiplex techniques evolved in recent years will provide new opportunities for detection of low molecular weight (LMW) proteome biomarker (peptides) by mass spectrometry. Improvements in peptidomics research were made based on separation of peptides and/or proteins by their physico-chemical properties in combination with mass spectrometric detection, respectively identification, and sophisticated bioinformatic tools for data analysis. To evaluate the potential of serological tumor marker detection by differential peptide display (DPD) we analyzed plasma samples from a tumor graft model. After subcutaneous injection of HCT-116 cells in immunodeficient mice and their growth to a palpable tumor, plasma samples were analyzed by DPD. The comparison of obtained mass spectrometric data allows discovery of tumor specific peptides which fit well into the biological context of cancer pathogenesis and show a strong correlation to tumor growth. The identified peptides indicate events associated with hyper-proliferation and dedifferentiation of cells from an epithelial origin, which are typical characteristics of human carcinomas. We conclude that these findings are a "proof of principle" to detect differentially expressed, tumor-related peptides in plasma of tumor-bearing mice.

Animals↗

PubMedDate not supplied

A high frequency electrical impedance tomograph using distributed parallel input channels.

Electrical Impedance Tomography (EIT) is an imaging technique based on multiple impedance measurements using electronically multiplexed surface electrodes. The present study describes an EIT prototype tomograph which uses two frequencies of applied signal: 31.25 kHz (within the frequency range originally used in early EIT studies) and 250 kHz. The use of the latter frequency was made possible as a result of several technical innovations including the use of separate groups of electrodes for current injection and for differential voltage measurement and the use of parallel input channels distributed around the object. In vitro images were successfully obtained using the system at the above two frequencies. It is concluded from this study that Electrical Impedance Tomography can be used at sufficiently high frequencies to enable bio-electrical tissue characterisation.

Electric Conductivity↗

PubMed2007

Polarization multiplexing and demultiplexing for appearance-based modeling.

Polarization has been used in numerous prior studies for separating diffuse and specular reflectance components, but in this work we show that it also can be used to separate surface reflectance contributions from individual light sources. Our approach is called polarization multiplexing and it has a significant impact in appearance modeling where the image as a function of illumination direction is needed. Multiple unknown light sources can illuminate the scene simultaneously, and the individual contributions to the overall surface reflectance are estimated. Polarization multiplexing relies on the relationship between the light source direction and the intensity modulation. Inverting this transformation enables the individual intensity contributions to be estimated. In addition to polarization multiplexing, we show that phase histograms from the intensity modulations can be used to estimate scene properties including the number of light sources.

Algorithms↗

PubMed2005

High-resolution Hadamard transform microscope fluorescence imaging: quantifying the DNA content in single cells.

This study presents a novel miniaturized Hadamard transform fluorescence imaging microscope, by combining a conventional fluorescence microscope with Hadamard transform multiplexing encoding using a one-dimensional movable mask to realize spatial resolution and a linear CCD for multichannel detection. The microscope can provide high-resolution automatically-generated 0-255 gray level HT images for morphological analysis and visualization of a single cell, and normalized HT images for cellular quantitative measurements. The microscope's imaging capability was applied to measure the DNA content in human lymphocyte, chicken erythrocyte and eel erythrocyte, and a comparative study was performed. The results show that the calibrated DNA content in a chicken erythrocyte is 2.32 pg when human lymphocyte is used as the standard, and eel erythrocyte may be a potentially reliable and novel standard for determining DNA contents in other species because it has a stable DNA value of 2.06 pg, with a CV of 4.3% when 20 eel erythrocytes are measured. The results also demonstrate that the HT imaging microscope should be valuable in the fields of medicine and cell biology.

DNA↗

PubMed2006

Chemical imaging by single pulse interferometric coherent anti-stokes Raman scattering microscopy.

A single pulse interferometric coherent anti-Stokes Raman (CARS) spectroscopy method is used to obtain broadband CARS spectra and microscopy images of liquid and polymer samples. The pump, Stokes, and probe pulses are all selected inside a single broadband ultrafast pulse by a phase- and polarization-controlled pulse shaping technique and used to generate two spectral interference CARS signals simultaneously. The normalized difference of these two signals provides an amplified background-free broadband resonant CARS spectrum over the 400-1500 cm(-1) range with 35 cm(-1) spectral resolution. Chemically selective microscopy images of multicomponent polymer and liquid samples are investigated with this new CARS method. Multiplex CARS spectra at 10,000 spatial points are measured within a few minutes, and used to construct chemically selective microscopy images with a spatial resolution of 400 nm. The spectral bandwidth limits, sensitivity, homodyne amplification advantages, spatial resolution, depolarization, chromatic aberration, and chemical imaging aspects of this new technique are discussed in detail.

Journal Article↗

PubMedDate not supplied

Classification of spectroscopically encoded resins by Raman mapping and infrared hyperspectral imaging.

Barcoded resins (BCRs) were recently introduced as a potential platform for pre-encoded multiplexed synthesis, screening, and biomedical diagnostics. A key step toward the development of this strategy is the ability to rapidly interrogate and classify the BCRs in a high-throughput, noninvasive manner. Here, we describe a one-step strategy based on Raman mapping and Fourier transform infrared imaging to classify and spatially resolve randomly distributed BCRs. To illustrate this methodology, mixtures of up to 25 different BCRs were imaged and classified with 100% confidence. This strategy can be readily extended to a larger pool of resins, provided each BCR features a unique vibrational fingerprint (spectroscopic barcode). We have also established that reliable single-bead Raman spectra can be recorded in 10 ms, thus confirming that Raman mapping, in particular, could be a very fast method to classify the BCRs.

Models, Molecular↗

PubMed2005

Integrated expressional analysis: application to the drug discovery process.

Microarray technology enables high-throughput testing of gene expression to investigate various neuroscience related questions. This in turn creates a demand for scalable methods to confirm microarray results and the opportunity to use this information to discover and test novel pathways and therapeutic applications. Discovery of new central nervous system (CNS) treatments requires a comprehensive understanding of multiple aspects including the biology of a target, the pathophysiology of a disease/disorder, and the selection of successful lead compounds as well as efficient biomarker and drug disposition strategies such as absorption (how a drug is absorbed), distribution (how a drug spreads through an organism), metabolism (chemical conversion of a drug, if any, and into which substances), and elimination (how is a drug eliminated) (ADME). Understanding of the toxicity is also of paramount importance. These approaches, in turn, require novel high-content integrative assay technologies that provide thorough information about changes in cell biology. To increase efficiency of profiling, characterization, and validation, we established a new screening strategy that combines high-content image-based testing on Array Scan (Cellomics) with a confocal system and the multiplexed TaqMan RT-PCR method for quantitative mRNA expression analysis. This approach could serve as an interface between high-throughput microarray testing and specific application of markers discovered in the course of a microarray experiment. Markers could pinpoint activation or inhibition of a molecular pathway related, for instance, to neuronal viability. We demonstrate the successful testing of the same cell population in an image-based translocational assay followed by poly(A) mRNA capture and multiplexed single tube RT-PCR. In addition, Ciphergen ProteinChip analysis can be performed on the supernatant, thus allowing significant complementarity in the data output and interpretation by also including the capture and initial analysis of proteins in the integrative approach presented. We have determined various conditions including the number of cells, RT and PCR optimization, which are necessary for successful detection and consequent assay integration. We also show the successful convergence of various different approaches and multiplexing of different targets within a single real-time PCR tube. This novel integrative technological approach has utility for CNS drug discovery, target and biomarker identification, selection and characterization as well as for the study of toxicity- and adverse event-associated molecular mechanisms.

Drug Evaluation, Preclinical↗

PubMed2003

Electrical impedance tomography spectroscopy (EITS) for human head imaging.

Electrical impedance tomography (EIT) is a recently developed medical imaging method which has practical advantages for imaging brain function as it is inexpensive, rapid and portable. Its principal use in validated human studies to date has been to image changes in impedance at a single excitation frequency over time, but there are potential applications where it is desirable to obtain images from a single point in time, which could be achieved by imaging over multiple frequencies. We describe a novel multifrequency EIT design which provides up to 64 electrodes for imaging in the head. This was achieved by adding a multiplexer to a single channel of an existing system, the Sheffield Mark 3.5. This provides a flexible protocol for addressing up to 64 electrodes but CMRR decreases from 90 dB to 80 dB and analogue amplifier bandwidth from > 1.6 MHz to 0.8 MHz. This did not significantly affect performance, as cylinders of banana, 10% of the diameter of a saline filled spherical tank, could be visualized with frequency referenced imaging. The design appears to have been an acceptable compromise between practicality and performance and will now be employed in clinical trials of multifrequency EIT in stroke, epilepsy and neonatal brain injury.

Electric Impedance↗

PubMed2006

Aberration properties in a chirped grating for coarse wavelength division demultiplexing.

We studied the imaging performance of a chirped grating for a demultiplexer designed for coarse wavelength division multiplexing using a wavefront aberration analysis and the ray tracing simulation. The demultiplexer was composed of a chirped grating, cylindrical lenses, and a waveguide. The best image point and the spot shape focused by the chirped grating were effectively calculated with the wavefront aberration. We applied the aberration analysis to design a waveguide to connect branched beams to photodetectors, and we confirmed the demultiplexing performance experimentally.

Journal Article↗

PubMed2002

Multiplexed computer-generated hologram with polygonal apertures.

A novel type of multiplexed computer-generated hologram (CGH) is designed with more than one billion of pixels per period. It consists of elementary cells divided into arbitrary-shaped polygonal apertures, the division being identical in all cells. The cells are further digitized into pixel arrays to exploit the huge space-bandwidth product of electron-beam lithography. The polygonal apertures in the same location inside the cells constitute a subhologram. With the Abbe transform that has never, to our knowledge, been used in other CGH designs, the subhologram images (subimages) are obtained with fast Fourier transforms. It is therefore possible to design a multiplexed CGH that has a size thousands of times larger than the manageable size of a conventional CGH designed with the iterative Fourier transform algorithm (IFTA). A much larger object window than that of the conventional CGH can also be achieved with the multiplexed polygonal-aperture CGH, owing to its extremely large dimensions. The multiplexed polygonal-aperture CGH is designed with the novel iterative subhologram design algorithm, which considers the coherent summation of the subimages and applies constraints on the total image, subimages, and subholograms. As a result, the noise appearing in the preceding multiplexed-CGH designs is avoided. The multiplexed polygonal-aperture CGH has a much higher diffraction efficiency than that resulting from either the preceding multiplexed-CGH designs or the conventional CGH designed by the IFTA.

Journal Article↗

PubMed2004

Cross talk between holograms of finite contrast in a phase-code multiplexing system.

The cross talk between holograms recorded in a phase-code multiplexing system is analyzed. The signal-to-noise ratio (SNR) is derived for the original image of a finite contrast ratio in the presence of a phase-shift error of the reference beam. It is shown that one particular code in the Walsh-Hadamard code set can be the dominant cross-talk noise source. We experimentally measured the SNR of the holograms recorded in an 8-bit phase-code multiplexing system by varying the contrast ratio of the original image. We show good agreement between experiment and theory. The existence of one bad code is also shown experimentally.

Journal Article↗

PubMed2006

Dynamic green fluorescent protein sensors for high-content analysis of the cell cycle.

We have developed two dynamic sensors that report cell cycle position in living mammalian cells. The sensors use well-characterized components from proteins that are spatially and temporally regulated through the cell cycle. Coupling of these components to Enhanced Green Fluorescent Protein (EGFP) has been used to engineer fusion proteins that report G1/S and G2/M transitions during the cell cycle without perturbing cell cycle progression. Expression of these sensors in stable cell lines allows high content analysis of the effects of drugs and gene knockdown on the cell cycle using automated image analysis to determine cell cycle position and to abstract correlative data from multiplexed sensors and morphological analysis.

Animals↗

PubMed2006

A rapid and simple system of detecting deletions on the Y chromosome related with male infertility using multiplex PCR.

Around 10% of males with idiopathic azoospermia or oligozoospermia, which are important causes of male infertility, have partial deletions on the long arm of the Y chromosome. To develop a rapid and accurate detection system for screening major Y deletions found in infertile men, we developed a multiplex PCR system that can simultaneously amplify five loci on the Y chromosome, SRY, AMELY, DBY, RBMY, DAZ and one locus on the X chromosome, AMELX. The size of the PCR products was designed to increase gradually from the distal Yp to the distal Yq. Our system could detect deletions of three major candidate regions for the azoospermic factor, AZFa, AZFb and AZFc on the Y chromosome together with sex. The gradual increase in the size of the PCR products was convenient for imaging the location of deletions on the Y chromosome. Moreover, the multiplex PCR system was combined with microchip-based electrophoresis, and the PCR products derived from each locus were separated within 4 min. Our system is useful for screening Y chromosomes bearing the structural anomalies including three major AZF deletions found among azoospermic or oligozoospermic males.

Base Sequence↗