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Results for “multiplex imaging”

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At least 109 records · Page 6Linked to original sources

PubMed2003

Filter multiplexing by use of spatial Code Division Multiple Access approach.

The increasing popularity of optical communication has also brought a demand for a broader bandwidth. The trend, naturally, was to implement methods from traditional electronic communication. One of the most effective traditional methods is Code Division Multiple Access. In this research, we suggest the use of this approach for spatial coding applied to images. The approach is to multiplex several filters into one plane while keeping their mutual orthogonality. It is shown that if the filters are limited by their bandwidth, the output of all the filters can be sampled in the original image resolution and fully recovered through an all-optical setup. The theoretical analysis of such a setup is verified in an experimental demonstration.

Journal Article↗

PubMed2006

Multiplex analysis of inflammatory signaling pathways using a high-content imaging system.

This chapter describes a robust high-content cellular screening assay to simultaneously analyze the spatiotemporal activation of three different kinase-associated signaling pathways involving NF-kappaB, JNK, and p38, all of which are closely implicated in proliferative and proinflammatory responses. Signal transduction is dependent on the translocation of NF-kappaB p65 and phosphorylated c-Jun and p38 from the cytosol to the nucleus, and fluorescent immunolabeling was used to monitor changes in their cellular distribution. Cellular screening, data acquisition, and data interpretation were conducted on the ArrayScan HCS Reader (Cellomics Inc., Pittsburgh, PA). Assay adaptation to various cellular systems is feasible when sufficient separation of the nuclear and cytosolic compartment can be achieved and if cell adhesion properties permit proper attachment to the culture plates. Substitution of NF-kappaB p65 and phosphorylated forms of c-Jun and p38 as targets to analyze other translocating components is possible and is limited primarily by antibody specificity and the risk of fluorescent bleed-through between emission channels. Because assay validity is particularly confounded by inadequate spectral separation of the detection dyes in multicolor labeling assays, means of eliminating or counterbalancing staining artifacts are illustrated. Also, protocol parameter settings important for imaging and image processing are described, including object identification, image exposure settings, separation of cytosolic and nuclear regions, number of cells sufficient for analysis, and the use of gating thresholds critical for cell sorting and subpopulation analysis. This assay is a useful tool to investigate the interplay between signaling pathways and the mode of action, potency, and selectivity of compound inhibition of specific target molecules in a cellular context.

Cell Nucleus↗

PubMed2007

The HaloTag: a novel technology for cell imaging and protein analysis.

The ability to specifically label proteins with a wide range of optical properties and functionalities can help reveal information about protein functions and dynamics in living cells. Here, we describe a technology for covalent tethering of organic probes directly to a specially designed reporting protein expressed in live cells. The reporting protein can be used in a manner similar to green fluorescent protein, except that the fluorophore might be interchanged among a variety of standard dyes. This allows living cells to be imaged at different wavelengths without requiring changes to the underlying genetic constructs, and the colors can be rapidly switched to allow temporal analysis of protein fate. The stability of the bond permits imaging of live cells during long time periods, imaging of fixed cells, and multiplexing with different cell/protein analysis techniques. The dyes can also be exchanged with other functional molecules, such as biotin to serve as an affinity handle, or even solid supports for direct covalent immobilization. The technology complements existing methods and provides new options for cell imaging and protein analysis.

Animals↗

PubMed2004

Volume holographic hyperspectral imaging.

A volume hologram has two degenerate Bragg-phase-matching dimensions and provides the capability of volume holographic imaging. We demonstrate two volume holographic imaging architectures and investigate their imaging resolution, aberration, and sensitivity. The first architecture uses the hologram directly as an objective imaging element where strong aberration is observed and confirmed by simulation. The second architecture uses an imaging lens and a transmission geometry hologram to achieve linear two-dimensional optical sectioning and imaging of a four-dimensional (spatial plus spectral dimensions) object hyperspace. Multiplexed holograms can achieve simultaneously three-dimensional imaging of an object without a scanning mechanism.

Journal Article↗

PubMed2002

Effective time averaging of multiplexed measurements: a critical analysis.

Multiplexing and time averaging of signal are effective noise reduction protocols applied in many analytical measurement systems. The efficacy of these protocols may be reduced by random occurrences of high-magnitude noise that do not conform to the statistical distribution of noise for all other measurements in the data set. This high-magnitude noise, which may have an insignificant probability of occurrence for a single measurement, almost certainly affects data collected in a multichannel, multiplexed modality, such as Fourier transform infrared (FT-IR) spectroscopic imaging employing focal plane array detectors. To recover time-averaging advantages in these cases, we present a general coaddition method that uses two statistical measures, the mean and median of the ensemble of measurements of a signal, to obtain a better estimate of the true signal than that estimated by time averaging alone. This method, termed median filtered time averaging, is shown to be an effective noise removal procedure for FT-IR imaging data. The effects of noise removal on time averaging and multiplexing are examined theoretically and are demonstrated for hyperspectral infrared microspectroscopic imaging data obtained from human skin biopsies by using a rapid data acquisition procedure.

Spectroscopy, Fourier Transform Infrared↗

PubMed1986

A multinuclear magnetic resonance imaging technique--simultaneous proton and sodium imaging.

Simultaneous imaging of proton and sodium is achieved using a new two coil system and a time-multiplexing technique with a 1.5 T NMR imaging system. Distinctly different NMR parameters of protons and sodium, such as resonant frequencies and T1 relaxation rates, are incorporated in the imaging with a new dual coil arrangement. Since the T1 relaxation time of sodium is substantially shorter than that of protons, a set of fast repeating sodium pulse sequences is inserted between the proton pulse repetition intervals. The different resonant frequencies of the nuclei provide good isolation between the two RF coils. In this paper, the system configuration and the pulse sequence employed for the proposed simultaneous multinuclear imaging of protons and sodium are presented along with some preliminary results.

Humans↗

PubMed2005

Difficult to diagnose manifestations of vasculitis: does an interdisciplinary approach help?

In the early stages of disease, primary systemic vasculitides often present with non-specific symptoms that make early diagnosis a challenge. The variety of clinical manifestations found in systemic vasculitis is huge, and some manifestations are frequently not clinically overt at first presentation. A logical implication of the often non-specific and sometimes subclinical presentation of vasculitis is that a systematic diagnostic work-up is necessary. This requires a multidisciplinary approach involving the expert opinion of specialists from many disciplines, such as neurology, radiology, respiratory medicine, pathology and microbiology. There are no generally accepted diagnostic criteria for primary systemic vasculitides, and the application of classification criteria as diagnostic criteria is not feasible and may even be misleading. The demonstration of vasculitis on biopsy is still the gold standard for the diagnosis of vasculitis. In cases where biopsies cannot be obtained, surrogate parameters of vasculitis (e.g. glomerular hematuria or mononeuritis multiplex), along with serology and imaging, can support a clinical diagnosis of vasculitis. This review discusses the approach to the diagnosis of central nervous system and pulmonary manifestations of primary systemic vasculitis. These two examples of difficult to diagnose manifestations of vasculitis illustrate the necessity of an interdisciplinary approach to the diagnostic work-up.

Diagnosis, Differential↗

PubMed2006

High-resolution, high-throughput microscopy analyses of nuclear receptor and coregulator function.

Steroid nuclear receptors are ligand-dependent transcription factors that have been studied since the early 1960s by principally biochemical and reporter assay approaches. From these studies an elegant and complex model of nuclear receptor transcription regulation has been developed. Inherent to both biochemical and reporter assay approaches is the generation of averaged responses and it is not generally considered that individual cells could exhibit quite varied responses. In some cases, recent microscopic single-cell analyses provide markedly different responses relative to traditional approaches based on population averaging and underscore the need to continue refinement of the current model of nuclear receptor-regulated transcription. While single-cell analyses of nuclear receptor action have been hindered by the predominantly qualitative nature of the approach, high-throughput microscopy is now available to resolve this issue. This chapter demonstrates the utility of high-throughput microscopic analyses of nuclear receptor and nuclear receptor coregulator function. The ability of high-throughput microscopy to generate physiologically appropriate test populations by filtering based on morphological and protein of interest expression criteria is demonstrated. High-resolution, high-throughput microscopy is illustrated that provides quantitative subcellular information for both androgen and estrogen receptors. Efforts are ongoing to develop model systems that provide additional multiplex data and with refined image analyses to achieve true high-content imaging screens.

Algorithms↗

PubMed2025

Impact of stromal maturity and proportion on prognosis and immune landscape in colorectal cancer.

BACKGROUND: Tumour microenvironment and cancer cells have constant interaction affecting cancer progression. Tumour-stroma ratio (TSR) in the tumour centre and desmoplastic reaction (DR) classification at the invasive margin are prognostic factors based on stroma evaluation on H&E slides. However, their combined value and immunological associations remain poorly defined. This study examines the prognostic and immunological value of TSR, DR, and their combination in two large colorectal cancer cohorts. METHODS: Two colorectal cancer cohorts (N = 1,876) were analyzed. We introduced a three-tiered Stromal Maturity and Proportion Score (SMAPS) based on the presence of high (>50%) TSR and myxoid stroma (immature DR classification). Alcian blue staining was used to further quantify myxoid stroma. Multiplex immunohistochemistry combined with digital image analyses, was utilized to study immune cell densities associated with SMAPS, TSR, DR, and Alcian blue intensity. RESULTS: In the study cohort (N = 1,100), SMAPS was a stronger predictor of cancer-specific mortality [HR for high (vs. low) SMAPS 2.01 (95% CI 1.47-2.75), p < 0.0001] compared to TSR [HR for stroma-high (vs. stroma-low) 1.49 (95% CI 1.15-1.93), p = 0.003] and DR classification [HR for immature (vs. mature) 1.84 (95% CI 1.39-2.45), p < 0.0001]. High SMAPS, stroma-high TSR, and immature DR correlated with lower densities of CD3+ T cells, B cells, M1-like macrophages, CD66B+ granulocytes, and mast cells. Alcian blue staining was associated with immature DR and corresponding immune cells. The validation cohort (N = 776) confirmed the association of SMAPS with survival and T cell densities. CONCLUSIONS: TSR and DR are independent prognostic factors for cancer-specific survival. SMAPS is a promising prognostic tool that integrates stromal maturity at the invasive margin and stromal proportion in the tumour centre. SMAPS has stronger prognostic value compared to TSR and DR classifications alone. A high stromal proportion and myxoid content are associated with an immunosuppressive microenvironment characterized by lower densities of antitumourigenic immune cells.

Humans↗

PubMed2026

Spatial Transcriptomics Identifies Characteristic Immunological Niches in Atopic Dermatitis.

BACKGROUND: Atopic dermatitis (AD) is primarily driven by a Type 2 immune response, with T helper (TH2) cells producing IL-4 and IL-13, thereby promoting inflammation, itch, and a compromised skin barrier. Yet, the spatial organization of pathogenic immune cells and their interactions with stromal and epithelial compartments in human AD skin remain incompletely understood. METHODS: We performed 10× Genomics Visium spatial transcriptomics on FFPE skin biopsies from patients with AD (n = 6), psoriasis (n = 2), and healthy controls (n = 5). Data were integrated with AD single-cell RNA sequencing (scRNA-seq) datasets and complemented by imaging mass cytometry (IMC) and multiplex immunofluorescence (IF) to validate the spatial localization of immune cells. Cell-cell communication analysis revealed putative signaling interactions within immune niches. RESULTS: Spatial clustering resolved tissue compartments and demonstrated transcriptional dysregulation in keratinocytes in AD and psoriasis. AD lesions showed a conserved spatial organization of immune aggregates within the superficial dermis. Integration of scRNA-seq signatures revealed spatially organized co-localization of T cells and mature migratory dendritic cells (mmDCs). We developed a ring-based neighborhood analysis to characterize the cellular organization of the immune-stromal niches, revealing T cell-enriched regions surrounded by inflammatory fibroblasts and activated keratinocytes. Intercellular communication analysis further identified putative signaling within mmDC-T cell niches that may promote pathogenic T cell recruitment and activation. Application of tertiary lymphoid structure (TLS) signatures indicated the presence of TLS-like regions. IMC and IF validated the close spatial proximity between activated TH2 cells and mmDCs. CONCLUSION: AD lesions contain spatially organized TLS-like immune niches at the dermal-epidermal junction, characterized by the close association of T cells and mmDCs and coordinated interactions with surrounding stromal and epithelial compartments. These mmDC-T cell niches may represent potential targets for future therapeutic strategies aimed at disrupting persistent local inflammatory pathways and improving long-term disease control.

atopic dermatitis↗

PubMed2003

Orthogonal aperture multiplexing for multilayered waveguide holographic read-only memories.

A multilayered waveguide holographic read-only memory is a promising candidate for the next generation of optical data storage systems. We improved the data density of the memory by using a multiplexing method with a set of orthogonal optical masks. We multiplexed as many as nine images into one waveguide hologram, and all the observed images had negligible cross talk. This made it possible to achieve a ninefold increase in data density. We provide experimental results for both metallic and liquid-crystal masks.

Journal Article↗

PubMed2004

Autostereoscopic three-dimensional display based on a micromirror array.

A novel approach for three-dimensional (3-D) display systems implemented with a micromirror array was proposed, designed, realized, and tested. The major advantages of this approach include the following: (1) micromirrors are reflective and hence achromatic (panchromatic), (2) a wide variety of displays can be used as image sources, and (3) time multiplexing can be introduced on top of space multiplexing to optimize the viewing zone arrangements. A two-view (left and right) 3-D autostereoscopic display system was first constructed. Left- and right-eye views in the forms of both still and motion 3-D scenes were displayed, and viewers were able to fuse the stereo information. A multiview (two left and two right) 3-D autostereoscopic display system was then simulated.

Journal Article↗

PubMed2006

Three-dimensional vibrational imaging of a microcrystalline J-aggregate using supercontinuum-based ultra-broadband multiplex coherent anti-stokes Raman scattering microscopy.

Vibrational properties of a porphyrin J-aggregate microcrystal have been investigated by ultra-broadband multiplex coherent anti-Stokes Raman scattering (CARS) microspectroscopy using a supercontinuum light source generated from a photonic crystal fiber. Owing to a strong resonance effect due to an excitonic transition, clear spectral and spatial profiles of the CARS signal have been successfully obtained. On the basis of the comparison between the CARS and the fluorescence images, the spatial dependence of the CARS signal can be explained by the spatial inhomogeneity of the excitonic transition energy in the single J-aggregate microcrystal.

Journal Article↗

PubMed1972

Multiplexing ultrasonic wave fronts by holoraphy.

Easily viewable, three-dimensional images have been produced from information derived from the passage of sound through the head of a living human subject. In this technique a new form of holographic multiplexing is used to construct the three-dimensional image from two-dimensional ultrasonic B-scans taken in many separated planes.

Head↗

PubMedDate not supplied

Neurogenic arthrogryposis multiplex congenita: clinical and MRI findings.

A clinical and magnetic resonance imaging (MRI) study on a selected group of 11 children, with a diagnosis of neurogenic arthrogryposis multiplex congenita (AMC) based on clinical, electromyographic, and muscle biopsy findings, is presented to determine the extent of central nervous system involvement in AMC. Family history, pregnancy, perinatal problems, other abnormalities, and epileptic seizures were reviewed. Neurologic examination, electroencephalography, intellectual assessment, and MRI study both of spinal cord and brain were performed. The clinical and laboratory findings disclosed evidence of spinal cord lesions with involvement of anterior horn cell function in all patients, and impairment of cerebral function in 5 patients. MRI revealed spinal cord atrophy in 3 patients, diffuse atrophy in 2 patients, and involved thoraco-lumbar segments in 1 patient. Cranial MRI studies demonstrated features of developmental brain abnormalities in 3 patients, cortical frontal atrophy in 2, and was normal in 4. In neurogenic AMC patients, MRI examination of the spinal cord and brain may help to clarify the pathogenesis of the disease and is helpful for prognostic and therapeutic purposes.

Adolescent↗

PubMedDate not supplied

Volumetric multiplexed transmission holography of the heart with echocardiographic data.

Three-dimensional imaging enhances delineation of cardiac anatomy and function. Currently, three-dimensional echocardiography involves complex rendering techniques to imply depth in a given image. Also, display of the final images on a video monitor neutralizes the volume information of the object. A hologram is a true, three-dimensional replica of the original object and does not entail complex data processing. Holography has not been used previously in cardiovascular imaging. In this experiment, 11 excised mammalian whole hearts and five isolated left ventricles were imaged in a water bath. Parallel, tomographic, echocardiographic images comprised the data source from which holograms were obtained by the technique of multiplexed holography. Holograms were mounted on a holographic film and viewed in a special viewing box. High-quality holograms were obtained from every data set. Excellent gray-scale discrimination allowed crisp visualization of cardiac structures and simulated diseases. Electronic sectioning and various projections optimized viewing of the three-dimensional anatomy in surgical orientations. Thus holograms can be produced from tomographic cardiac ultrasound data and could open a new avenue in three-dimensional echocardiography.

Animals↗

PubMed2001

A sixteen-channel multiplexing upgrade for single channel receivers.

With the increasing interest in phased arrays in magnetic resonance imaging, imaging system receivers capable of acquiring larger number of parallel signals are needed. Suggested techniques for rapid imaging propose the use of arrays with as many as 128 elements. While simply duplicating the number of receiver chains as needed is a viable technique, it quickly becomes both cumbersome and expensive. Time domain multiplexing offers an alternative solution to this problem. By using RF multiplexing switches, a single receiver can be upgraded to an array receiver capable of multi-channel data acquisition giving users array capability. Additionally, it can be used to dramatically increase acquisition capability of multiple receiver systems. This paper reports results from a multiplexing system upgrade, which converts a single channel standard clinical imaging system to a 16-channel array system. The upgrade includes both the RF multiplexing front-end and an external data acquisition system with image processing capability. Issues concerning the implementation of high channel-count multiplexers are also discussed.

Equipment Design↗

PubMed2006

Surface plasmon resonance imaging measurements of protein interactions with biopolymer microarrays.

The surface-sensitive optical technique of surface plasmon resonance (SPR) imaging is an ideal method for the study of affinity binding interactions of unlabeled biological molecules in a multiplexed format. This approach has been widely applied to monitor DNA-DNA, DNA-RNA, peptide-protein, and protein-protein interactions as well as surface enzyme reactions. The success of SPR imaging measurements relies on the robust attachment of biomolecules in an array format. In this chapter, we introduce two different surface attachment chemistries that covalently immobilize DNA and peptides onto gold surfaces through the modification of self-assembled alkanethiol monolayers. Array fabrication approaches for the creation of individually addressable elements through the use of either gold dot patterns or polydimethylsiloxane (PDMS) microchannels are detailed. The utility of SPR imaging for the study of protein interactions is demonstrated with two biological systems: the binding of response regulator proteins, VanR and OmpR, onto a DNA array, and the interaction of S protein with an array of S-peptide variants. Furthermore, the application of real-time SPR imaging to the multiplexed determination of S-protein adsorption/desorption kinetics is described.

Bacterial Proteins↗