Characterization of color cross-talk of CCD detectors and its influence in multispectral quantitative phase imaging

Multi-spectral quantitative phase imaging (QPI) is an emerging imaging modality for wavelength dependent studies of several biological and industrial specimens. Simultaneous multi-spectral QPI is generally performed with color CCD cameras. However, color CCD cameras are suffered from the color crosstalk issue, which needed to be explored. Here, we present a new approach for accurately measuring the color crosstalk of 2D area detectors, without needing prior information about camera specifications. Color crosstalk of two different cameras commonly used in QPI, single chip CCD (1-CCD) and three chip CCD (3-CCD), is systematically studied and compared using compact interference microscopy. The influence of color crosstalk on the fringe width and the visibility of the monochromatic constituents corresponding to three color channels of white light interferogram are studied both through simulations and experiments. It is observed that presence of color crosstalk changes the fringe width and visibility over the imaging field of view. This leads to an unwanted non-uniform background error in the multi-spectral phase imaging of the specimens. It is demonstrated that the color crosstalk of the detector is the key limiting factor for phase measurement accuracy of simultaneous multi-spectral QPI systems.Comment: 16 pages, 8 figure

Multispectral fingerprinting for improved in vivo cell dynamics analysis

Background: Tracing cell dynamics in the embryo becomes tremendously difficult when cell trajectories cross in space and time and tissue density obscure individual cell borders. Here, we used the chick neural crest (NC) as a model to test multicolor cell labeling and multispectral confocal imaging strategies to overcome these roadblocks. Results: We found that multicolor nuclear cell labeling and multispectral imaging led to improved resolution of in vivo NC cell identification by providing a unique spectral identity for each cell. NC cell spectral identity allowed for more accurate cell tracking and was consistent during short term time-lapse imaging sessions. Computer model simulations predicted significantly better object counting for increasing cell densities in 3-color compared to 1-color nuclear cell labeling. To better resolve cell contacts, we show that a combination of 2-color membrane and 1-color nuclear cell labeling dramatically improved the semi-automated analysis of NC cell interactions, yet preserved the ability to track cell movements. We also found channel versus lambda scanning of multicolor labeled embryos significantly reduced the time and effort of image acquisition and analysis of large 3D volume data sets. Conclusions: Our results reveal that multicolor cell labeling and multispectral imaging provide a cellular fingerprint that may uniquely determine a cell's position within the embryo. Together, these methods offer a spectral toolbox to resolve in vivo cell dynamics in unprecedented detail

Lunar maria and related deposits: Preliminary Galileo imaging results

During the Earth-Moon flyby the Galileo Solid State Imaging system obtained new information on lunar media. Imaging data in spectral bands from 0.4 to 1.0 micron wavelength provide color data for deposits on the western limb. General objectives were to determine the composition and stratigraphy of mare and related deposits for areas not previously seen well in color, and to compare the results with well-studied nearside maria. Initial results from images reduced with preliminary calibrations show that Galileo spectral reflectance data are consistent with previous earthbased observations

Hinode/EIS spectroscopic validation of very hot plasma imaged with Solar Dynamics Observatory in non-flaring active region cores

We use coronal imaging observations with SDO/AIA, and Hinode/EIS spectral data, to explore the potential of narrow band EUV imaging data for diagnosing the presence of hot (T >~5MK) coronal plasma in active regions. We analyze observations of two active regions (AR 11281, AR 11289) with simultaneous AIA imaging, and EIS spectral data, including the CaXVII line (at 192.8A) which is one of the few lines in the EIS spectral bands sensitive to hot coronal plasma even outside flares. After careful coalignment of the imaging and spectral data, we compare the morphology in a 3 color image combining the 171, 335, and 94A AIA spectral bands, with the image obtained for CaXVII emission from the analysis of EIS spectra. We find that in the selected active regions the CaXVII emission is strong only in very limited areas, showing striking similarities with the features bright in the 94A (and 335A) AIA channels and weak in the 171A band. We conclude that AIA imaging observations of the solar corona can be used to track hot plasma (6-8MK), and so to study its spatial variability and temporal evolution at high spatial and temporal resolution.Comment: 10 pages, 2 figures, accepted for publication on ApJ Letter

Simple method for sub-diffraction resolution imaging of cellular structures on standard confocal microscopes by three-photon absorption of quantum dots

This study describes a simple technique that improves a recently developed 3D sub-diffraction imaging method based on three-photon absorption of commercially available quantum dots. The method combines imaging of biological samples via tri-exciton generation in quantum dots with deconvolution and spectral multiplexing, resulting in a novel approach for multi-color imaging of even thick biological samples at a 1.4 to 1.9-fold better spatial resolution. This approach is realized on a conventional confocal microscope equipped with standard continuous-wave lasers. We demonstrate the potential of multi-color tri-exciton imaging of quantum dots combined with deconvolution on viral vesicles in lentivirally transduced cells as well as intermediate filaments in three-dimensional clusters of mouse-derived neural stem cells (neurospheres) and dense microtubuli arrays in myotubes formed by stacks of differentiated C2C12 myoblasts

Navigating the roadblocks to spectral color reproduction: data-efficient multi-channel imaging and spectral color management

Commercialization of spectral imaging for color reproduction will require the identification and traversal of roadblocks to its success. Among the drawbacks associated with spectral reproduction is a tremendous increase in data capture bandwidth and processing throughput. Methods are proposed for attenuating these increases with data-efficient methods based on adaptive multi-channel visible-spectrum capture and with low-dimensional approaches to spectral color management. First, concepts of adaptive spectral capture are explored. Current spectral imaging approaches require tens of camera channels although previous research has shown that five to nine channels can be sufficient for scenes limited to pre-characterized spectra. New camera systems are proposed and evaluated that incorporate adaptive features reducing capture demands to a similar few channels with the advantage that a priori information about expected scenes is not needed at the time of system design. Second, proposals are made to address problems arising from the significant increase in dimensionality within the image processing stage of a spectral image workflow. An Interim Connection Space (ICS) is proposed as a reduced dimensionality bottleneck in the processing workflow allowing support of spectral color management. In combination these investigations into data-efficient approaches improve two critical points in the spectral reproduction workflow: capture and processing. The progress reported here should help the color reproduction community appreciate that the route to data-efficient multi-channel visible spectrum imaging is passable and can be considered for many imaging modalities

Using the Matrix R method for spectral image archives

Conventional color digital cameras can only produce three-channel images so they are limited when high-quality color reproduction is required. Alternatively, spectral imaging increases the number of channels and can retrieve spectral reflectance for each scene pixel. The major goal of spectral imaging is high spectral accuracy, while it may also be beneficial to achieve high colorimetric accuracy for a specific viewing condition. A new spectral reconstruction method, called the matrix R method, was developed to achieve both goals simultaneously. An experiment was performed to test this method. The experimental results have been very promising; average color difference for all targets evaluated was about 1.3 CIEDE2000 and 2.0% RMS. These results suggest that this new method is a promising method for building digital image databases for museums, archives and libraries

Lippman 2000: a spectral image database under construction

In support of research projects both within the Munsell Color Science Laboratory and outside which rely on having full knowledge of the spectral makeup of scenes, a number of methods for capturing spectral images are being explored. This project is named Lippmann2000 in honor of Gabriel Lippmann who in 1891 devised a method to perfectly reconstruct the spectral content of real world scenes. In spite of Lippmann’s invention, a more primitive three-channel model, first demonstrated by James Clerk Maxwell 30 years prior, has dominated the color imaging field. The Maxwellian model, universal in today’s silver halide and electronic color image capture systems, relies on the metameric properties of the human visual system to simulate the appearance of an original color. It has been recognized by those in the forefront of imaging research that the capture of full spectral data holds advantage over traditional three-channel methods. This paper describes our efforts to-date to build our database of spectral images

Multi-Color Imaging of Magnetic Co/Pt Multilayers

We demonstrate for the first time the realization of a spatial resolved two color, element-specific imaging experiment at the free-electron laser facility FERMI. Coherent imaging using Fourier transform holography was used to achieve direct real space access to the nanometer length scale of magnetic domains of Co/Pt heterostructures via the element-specific magnetic dichroism in the extreme ultraviolet spectral range. As a first step to implement this technique for studies of ultrafast phenomena we present the spatially resolved response of magnetic domains upon femtosecond laser excitation