Cooled Charge-coupled Device Research Articles

In vivo molecular imaging of adenoviral versus lentiviral gene therapy in two bone formation models

Regional gene therapy techniques are promising methods to enhance bone formation in large bone defects that would be difficult to treat with allograft or autograft bone stock. In this study, we compared in vivo temporal expression patterns of adenoviral- and lentiviral-mediated gene therapy in two bone formation models. Primary rat bone marrow cells (RBMC) were transduced with lentiviral or adenoviral vectors containing luciferase (Luc) or BMP-2 cDNA, or cotransduced with vectors containing Luc and bone morphogenetic protein 2 (BMP-2). In vitro protein production was determined with luciferase assay or ELISA (for BMP-2 production) weekly for 12 weeks. Two bone formation models were used -- a hind limb muscle pouch or radial defect -- in SCID mice. A cooled charged-coupled device (CCD) camera was used to image in vivo luciferase expression weekly for 12 weeks. In vitro, adenoviral expression of BMP-2 and luciferase was detected by ELISA or luciferase assay, respectively, for 4 weeks. Lentiviral expression of BMP-2 and luciferase was sustained in culture for 3 months. Using the CCD camera, we found that adenoviral vectors expressed luciferase expression for up to 21 days, but lentiviral vectors expressed target gene expression for 3 months in vivo in both bone formation models. There was no detectable difference in the amount of bone formed between the adenoviral and lentiviral groups. Lentiviral-mediated delivery of BMP-2 can induce long term in vitro and in vivo gene expression, which may be beneficial when developing tissue engineering strategies to heal large bone defects or defects with a compromised biologic environment.

Development of a “Built-in” Scanning Near Field Microscope Head for an Atomic Force Microscope System and Stress Mapping of an Al2O3/ZrO2 Eutectic Composite

We constructed a scanning near-field optical microscope (SNOM) on a commercially available atomic force microscopy (AFM) apparatus (SPM-9500J2; Shimadzu Corp.) to measure the stress distribution in ceramic composite materials. Features of our SNOM system are: (1) a compact SNOM head substituted for the original AFM head; (2) a wide scanning range (125 × 125 μm2) inherited from the original scanner; (3) use of conventional shear-force regulation; (4) an optical system for the illumination-collection (I-C) mode; (5) excitation by a 488 nm line of an Ar-ion laser, and (6) light detection by photon counting or a polychromator equipped with an electronically cooled charge coupled device (CCD). This SNOM system was used to measure the surface structure and stress distribution of an Al2O3/ZrO2 eutectic composite. We simultaneously measured topographic images and fluorescence spectra of an Al2O3/ZrO2 eutectic composite. We estimated its peak intensity, peak position, and peak width from the fluorescence spectrum during scanning, which respectively correspond to the abundance of Al2O3, stress in the grain, and the anisotropy of that stress. Mapping images showed that the stress and its anisotropy were weaker in the center of the Al2O3 grain than its boundary between Al2O3 and ZrO2. That observation suggests that Al2O3 underwent intense anisotropic stress induced by volume expansion in the phase transition of ZrO2 from the cubic phase to the monoclinic phase during preparation.

In vivo bioluminescence imaging of cord blood derived mesenchymal stem cell transplantation into rat myocardium

The conventional method for the analysis of myocardial cell transplantation depends on postmortem histology. Here, we have sought to demonstrate the feasibility of a longitudinal monitoring of transplanted cell survival in living animals, accomplished with optical imaging techniques and pharmacological interventions. Human cord blood (50 ml) was donated with parental consent. After getting cord blood derived mesenchymal stem cells (CBMSCs), cells were transfected (MOI = 100) overnight with adenovirus encoding firefly luciferase gene (Ad-CMV-Fluc). Our experimental Sprague-Dawley rats (n = 12) were given intramyocardial injections containing 1 x 10(6) CBMSCs, which had been made to express the firefly luciferase (Fluc) reporter gene. Optical bioluminescence imaging was then conducted using a cooled charged-coupled device (CCD) camera (Xenogen), beginning on the day after the transplantation (day 1). Groups of mice were intraperitoneally injected with cyclosporine (5 mg/kg) or tacrolimus (1 mg/kg), in an attempt to determine the degree to which cell survival had been prolonged, and these values were then compared with the cell survival values of the negative control group. The presence of transplanted CBMSCs on in vivo images confirmed by in situ hybridization for human specific Alu in the myocardium. Cardiac bioluminescence signals were determined to be present for 6 days after transplantation: day 1 (97000 +/- 9100 x 10(5) p/s/cm2/sr), day 3 (9600 +/- 1110 p/s/cm2/sr), and day 5 (3200 +/- 550 p/s/cm2/sr). The six mice that received either cyclosporine or tacrolimus displayed cardiac bioluminescence signals for a period of 8 days after transplantation. We observed significant differences between the treated group and the non-treated group, beginning on day 3 (tacrolimus; 26500 +/- 4340 p/s/cm2/sr, cyclosporine; 27200 +/- 3340 p/s/cm2/sr, non-treated; 9630 +/- 1180 p/s/cm2/sr, p < 0.01), and persisting until day 7 (tacrolimus; 12500 +/- 2946 p/s/cm2/sr, cyclosporine; 7310 +/- 1258 p/s/cm2/sr, non-treated; 2460 +/- 160 p/s/cm2/sr, p < 0.01). The human-derived CBMSCs were detected in the myocardium 3 days after transplantation by in situ hybridization. The locations, magnitude, and survival duration of the CBMSCs were noninvasively monitored with a bioluminescence optical imaging system. We determined that optical molecular imaging expedites the fast throughput screening of pharmaceutical agents, allowing for the noninvasive tracking of cell survival within animals. In rat cardiac CBMSC transplant models, transient immunosuppressive treatment with tacrolimus or cyclosporine was shown to improve donor cell survival.

Detection and Quantification of Protein Biomarkers from Fewer than 10 Cells

The use of antibody microarrays continues to grow rapidly due to the recent advances in proteomics and automation and the opportunity this combination creates for high throughput multiplexed analysis of protein biomarkers. However, a primary limitation of this technology is the lack of PCR-like amplification methods for proteins. Therefore, to realize the full potential of array-based protein biomarker screening it is necessary to construct assays that can detect and quantify protein biomarkers with very high sensitivity, in the femtomolar range, and from limited sample quantities. We describe here the construction of ultramicroarrays, combining the advantages of microarraying including multiplexing capabilities, higher throughput, and cost savings with the ability to screen very small sample volumes. Antibody ultramicroarrays for the detection of interleukin-6 and prostate-specific antigen (PSA), a widely used biomarker for prostate cancer screening, were constructed. These ultramicroarrays were found to have a high specificity and sensitivity with detection levels using purified proteins in the attomole range. Using these ultramicroarrays, we were able to detect PSA secreted from 100 LNCaP cells in 3 h and from just four LNCaP cells in 24 h. Cellular PSA could also be detected from the lysate of an average of just six cells. This strategy should enable proteomic analysis of materials that are available in very limited quantities such as those collected by laser capture microdissection, neonatal biopsy microspecimens, and forensic samples.

Phase resolved electroluminescence measurements in thin films of low density polyethylene using a charge coupled device camera

Electroluminescence (EL) produced by a commercially available additive free low density polyethylene film has been investigated under a 50 Hz AC electrical stress. The spatial distribution, spectral characteristics and the phase angle relationship of EL with respect to the 50 Hz applied AC voltage were studied using a sensitive Peltier cooled charge-couple device (CCD) camera. The experimental results from several test samples show that the EL can be divided into two different groups according to the EL spatial distribution: (1) homogeneous EL and (2) inhomogeneous EL. At the same applied voltage, the spectra of the homogenous and inhomogeneous EL were found to be significantly different. Homogeneous EL had maximum emission intensity in the red and near infrared (>700 nm), whereas the inhomogeneous emission had maximum intensity at shorter wavelengths (550-600 nm). Phase resolved measurements showed that homogeneous EL always produced equal emission intensity in the positive and negative half-cycles (symmetric distribution), whereas, inhomogeneous EL produced an asymmetric intensity distribution. To explain these results, it is suggested that homogeneous emission is associated with charge injection and bipolar recombination in the surface states on both sides of the sample, whereas inhomogeneous emission is associated with field enhancement at defects on one of the two sides of the film which allow charge to be injected into states more representative of the bulk material.

Convective Ionospheric Storms: A Major Space Weather Problem

Convective Ionospheric Storms: A Major Space Weather Problem

Resonance Light Scattering Imaging Detection of Single Suprahelical Species of DNA Induced by Porphine‐5,10,15,20‐tetrakis(p‐phenyltrimethylaminium)

AbstractA resonance light scattering (RLS) imaging method was proposed based on imaging and measuring the RLS features of single suprahelical species of DNA, and its application to DNA assay was also investigated. In acidic medium, porphine‐5,10,15,20‐tetrakis(p‐phenyltrimethylaminium) (PTPTMA), could stack along the molecular surface of DNA with the mode of long‐range assembly to induce the formation of suprahelical species of DNA, resulting in strong RLS signals in the range of 450–510 nm. Under the excitation of 488 nm light beam of argon ion laser source, single suprahelical species could be observed with the aid of a common microscope due to the strong scattered light emitted by the suprahelical species. By capturing the RLS images of the single suprahelical species with a cooled charge coupled device (CCD) camera, and analyzing the RLS data, herein an RLS imaging method of DNA was proposed based on the linear relationship between the counts of suprahelical species in the detection focus plane and the concentration of DNA in nanograms. When 1.8??µmol/L PTPTMA was employed, both calf thymus DNA (ct DNA) and fish sperm DNA (fs DNA) in the range of 25–1100 ng/mL could be detected with the limits of detection lower than 25 ng/mL (3σ). Four synthetic samples were detected satisfactorily with relative standard deviations less than 5.1%.

A photographic technique for assessing the viewing-angle performance of liquid-crystal displays

— Liquid-crystal displays (LCDs) have notable variation in luminance and perceived contrast as a function of the angle from which they are viewed. Though this is an important performance issue for LCDs, most evaluation techniques for assessing this variation have been limited to laboratory settings. This study demonstrates the use of a photographic technique for such an evaluation. The technique is based on an actively cooled charge-coupled-device (CCD) detector in combination with a macro lens covering a circular angular range (θ) of ±42.5°. The camera was used to evaluate the luminance and perceived contrast properties of an LCD. Uniform field images corresponding to 17 equally spaced gray-scale values in the digital driving level (DDL) range of the display system were acquired. The 12-bit gray-scale digital images produced by the camera were converted to luminance units (cd/m2) via the measured luminance vs. DDL response function of the camera. The changes in perceived contrast as a function of viewing angle were derived from the Barten model of the gray-scale response of the human-visual system using the methods proposed by the AAPM TG18 Report. The results of this photographic technique were compared to measurements acquired from a similar display using a Fourier-optics-based luminance meter. The results of the two methods generally agreed to within 5%. The photographic methods used were found to be accurate and robust for in-field assessment of the angular response of LCDs over the FOV of the camera.

Noninvasive imaging of protein‐protein interactions from live cells and living subjects using bioluminescence resonance energy transfer

This study demonstrates a significant advancement of imaging of a distance-dependent physical process, known as the bioluminescent resonance energy transfer (BRET2) signal in living subjects, by using a cooled charge-coupled device (CCD) camera. A CCD camera-based spectral imaging strategy enables simultaneous visualization and quantitation of BRET signal from live cells and cells implanted in living mice. We used the BRET2 system, which utilizes Renilla luciferase (hRluc) protein and its substrate DeepBlueC (DBC) as an energy donor and a mutant green fluorescent protein (GFP2) as the acceptor. To accomplish this objective in this proof-of-principle study, the donor and acceptor proteins were fused to FKBP12 and FRB, respectively, which are known to interact only in the presence of the small molecule mediator rapamycin. Mammalian cells expressing these fusion constructs were imaged using a cooled-CCD camera either directly from culture dishes or by implanting them into mice. By comparing the emission photon yields in the presence and absence of rapamycin, the specific BRET signal was determined. The CCD imaging approach of BRET signal is particularly appealing due to its capacity to seamlessly bridge the gap between in vitro and in vivo studies. This work validates BRET as a powerful tool for interrogating and observing protein-protein interactions directly at limited depths in living mice.

Polar Distribution Dynamics of Con A Binding Sites in Embryo Sacs is Temporally Coupled by the Fertilization Process

The binding site distribution of concanavalin agglutinin (Con A) and wheat germ agglutinin (WGA) on embryo sacs at various developmental stages of Torenia fournieri L was studied by using a cooled Charge Coupled Device (CCD) and fluorescent Con A and WGA probes. The distribution patterns of Con A and WGA binding sites on embryo sacs changed during the fertilization process. The fluorescent signal indicating Con A binding sites was distributed evenly on the surface of the embryo sac wall before anthesis, was much denser on the micropylar end of the embryo sac wall and looked like a corona on the day of anthesis. After pollination, stronger fluorescence was present on the micropylar end of the embryo sac wall and the filiform apparatus (FA), showing an obvious polar distribution. When the pollen tube entered the embryo sac and reached a synergid, the fluorescence was still concentrated on the micropylar end and FA, and started to appear on the synergid. After fertilization, the polar distribution of the fluorescence gradually disappeared and an even distribution pattern was observed again on the embryo sac wall. These results revealed that the dynamic distribution of Con A binding sites was temporally coupled with the process of fertilization. WGA binding site distribution on the embryo sac was also investigated and showed a simple pattern but also regularly changed during the process of fertilization. The variation of these lectin binding sites during the fertilization process suggests that lectin binding site interactions may play a role in the process.

Multimodality Radionuclide, Fluorescence, and Bioluminescence Small-Animal Imaging

The imaging of molecular events in the complex physiological interplay between organelles, cells, tissues, and organs in the whole organism is now more practical through the use of small-animal imaging technologies. Radionuclide based molecular imaging using single photon emission computed tomography (SPECT) and positron emission tomography (PET) scanners have been used for imaging intracellular enzymes, receptors, transporters, reporter gene expression, and cell trafficking in small animals such as mice and rats. Sensitive cooled charge-coupled device (CCD) cameras which detect emitted light from fluorescent and bioluminescent probes within an organism have been useful in imaging pathologic changes in diseases such as cancer and infectious disease in small-animal models. Each molecular imaging modality has differing strengths and weaknesses. However, the ability to generate molecular reporter strategies that can be applied in multiple detection systems will likely provide more robust imaging data sets than from any single modality. The ultimate function of multimodality reporter strategies will be to provide information about molecular targets in a continuum of research subjects from cells to small animals to human patients.

Liver-Targeted Gene Therapy by SV40-Based Vectors Using the Hydrodynamic Injection Method

Efficient reconstitution of defective genes in hepatocytes could be used to treat various liver and systemic diseases through gene therapy. To explore the potential of SV40-based vectors in liver gene therapy, we constructed SV/luc, an SV40 T-antigen replacement transduction vector, that was propagated on COS and COT cells, which supply the SV40 T-antigen in trans. For liver targeting, BALB/C mice were injected via the tail vein with SV/luc stocks containing 3 x 10(6) to 10(8) transducing units in a volume of 1-2 ml. Luciferase activity was monitored with a light-detection cooled charged-coupled device (CCCD) camera, which enables continuous in vivo measurement of luc expression. The SV40 vector proved to be efficient in gene delivery to the liver, leading to long-term (> or =107 days) transgene expression in hepatocytes. Optimal results were obtained with 3 x 10(6) to 3 x 10(7) transducing units. The hydrodynamic vector delivery method caused transient liver inflammatory changes, with full recovery within days. Low levels of SV40-neutralizing antibodies were detected in the sera of treated mice; however, there was no indication of vector or transgene-specific cellular immune responses. Vectors packaged in vitro, using recombinant capsid proteins and plasmid DNA, were also effective in liver transduction. These results suggest that SV40 vectors may be useful for liver gene therapy.

Probing lectin binding sites on isolated viable generative and sperm cells of tobacco

Generative cells and sperm cells of tobacco were isolated and labeled by fluorescein isothiocyanate conjugated concanavalin agglutinin (FITC–Con A), wheat germ agglutinin (FITC–WGA) and soybean agglutinin (FITC–SBA) to compare lectin binding site (LBS) distributive patterns between generative cells and sperm cells, and between a pair of sperm cells from the same pollen tube. With the help of cooled charge couple device (CCD) and confocal microscope, Con A binding sites were found on the surface of 96% generative cells and 91% sperm cells. WGA binding sites were also detected on plasma membrane of generative cells and inner plasma membrane of vegetative cells and the crosswall of male germ unit (MGU), but it was hardly found on most of sperm cell surface. No soybean agglutinin (SBA) binding sites were detected on the plasma membrane of both generative cells and sperm cells. The experiment revealed that LBSs on the membrane surface of germ cell were reorganized or modified during the development from generative cell to sperm cell, indicating sperm formation might couple with its function specialization. The possible dimorphism on surface glycoprotein distribution of sperm cells was not confirmed in tobacco in this experiment since sperm cells derived from the same generative cell could be both labeled or not labeled at all by FITC–Con A. Based on the finding in present work and our former reports, we proposed that there might be functional difference of sperm in the population of living sperms of higher plants.

Emission spectra of bioluminescent reporters and interaction with mammalian tissue determine the sensitivity of detection in vivo

In vivo bioluminescence imaging depends on light emitted by luciferases in the body overcoming the effect of tissue attenuation. Understanding this relationship is essential for detection and quantification of signal. We have studied four codon optimized luciferases with different emission spectra, including enzymes from firefly (FLuc), click beetle (CBGr68, CBRed) and Renilla reniformins (hRLuc). At 25 degrees C, the in vitro lambda(max) of these reporters are 578, 543, 615, and 480 nm, respectively; at body temperature, 37 degrees C, the brightness increases and the firefly enzyme demonstrates a 34-nm spectral red shift. Spectral shifts and attenuation due to tissue effects were evaluated using a series of 20-nm bandpass filters and a cooled charge-coupled device (CCD) camera. Attenuation increased and the spectra of emitted light was red shifted for signals originating from deeper within the body relative to superficial origins. The tissue attenuation of signals from CBGr68 and hRLuc was greater than from those of Fluc and CBRed. To further probe tissue effects, broad spectral emitters were created through gene fusions between CBGr68 and CBRed. These resulted in enzymes with broader emission spectra, featuring two peaks whose intensities are differentially affected by temperature and tissue depth. These spectral measurement data allow for improved understanding of how these reporters can be used in vivo and what they can reveal about biological processes in living subjects.

Development of neutron radioscopy for the inspection of CF188 flight control surfaces

Neutron radioscopy, using a cooled charged coupled device (CCD) camera and a neutron-sensitive scintillation screen, was developed at the SLOWPOKE-2 Facility at the Royal Military College (RMC) to detect water ingress into the composite layers and the aluminium honeycomb core in flight control surfaces on the CF188 Hornet aircraft. The response of the CCD camera system was tested at different neutron fluxes utilising the SLOWPOKE-2 at RMC and the Breazeale Nuclear Reactor at Pennsylvania State University.

Deep-Ultraviolet Raman Microspectroscopy: Characterization of Wide-Gap Semiconductors

We have developed a high-throughput deep-ultraviolet (DUV) Raman microspectrometer with excitation from a continuous wave (cw) laser operated at 244 nm that enables us to characterize thin surface layers of wide-gap semiconductors. This spectrometer system consists of a filter spectrometer for the rejection of stray light and a high-dispersion spectrograph combined with a liquid nitrogen cooled charge-coupled device (CCD) detector and extends the low-frequency limit of the observable spectral range down to 170 cm(-1). In the microscope we use a Cassegrain reflective objective for the collection of the scattered light and an off-axis mirror for introduction of the excitation laser light. DUV Raman spectroscopy has been applied for studying wide-gap semiconductors including SiC and AlGaN epitaxial films and shallow implanted layers of these materials. Raman spectra of various crystals have also been measured for examining the performance of this system. Resonance enhancement of Raman bands has been observed for several semiconductors, and the results are discussed.

Use of a highly sensitive two-dimensional luminescence imaging system to monitor endogenous bioluminescence in plant leaves

BackgroundAll living organisms emit spontaneous low-level bioluminescence, which can be increased in response to stress. Methods for imaging this ultra-weak luminescence have previously been limited by the sensitivity of the detection systems used.ResultsWe developed a novel configuration of a cooled charge-coupled device (CCD) for 2-dimensional imaging of light emission from biological material. In this study, we imaged photon emission from plant leaves. The equipment allowed short integration times for image acquisition, providing high resolution spatial and temporal information on bioluminescence. We were able to carry out time course imaging of both delayed chlorophyll fluorescence from whole leaves, and of low level wound-induced luminescence that we showed to be localised to sites of tissue damage. We found that wound-induced luminescence was chlorophyll-dependent and was enhanced at higher temperatures.ConclusionsThe data gathered on plant bioluminescence illustrate that the equipment described here represents an improvement in 2-dimensional luminescence imaging technology. Using this system, we identify chlorophyll as the origin of wound-induced luminescence from leaves.

Optical imaging of transferrin targeted PEI/DNA complexes in living subjects.

Noninvasive optical bioluminescence imaging systems are important tools for evaluating gene expression in vivo for study of individual and temporal variation in a living animal. In this report, we demonstrate that expression of the firefly luciferase reporter gene (fl) delivered by transferrin (Tf) targeted polyethylenimine (PEI) complexes with, or without, poly(ethylene glycol) (PEG) modifications can be imaged in living A/J mice bearing N2A tumors using a cooled charged coupled device (CCD) camera. Tf-PEI-PEG, Tf-PEI, and PEI (positive control) complexes were tail-vein injected and mice were imaged at 5, 24, 48, and 72 h after complex injection. After imaging, the organs were analyzed ex vivo for firefly luciferase protein (FL) activity. The Tf and PEG modified formulations show significantly (P<0.05) higher FL activity in vivo and ex vivo at the tumor as compared to other organs, including the lungs (a site of high expression with PEI, the positive control). Furthermore, the in vivo bioluminescent signal correlated well (R(2)=0.83) with ex vivo FL activity. These data support that noninvasive imaging of fl reporter expression can be used to monitor the specificity of Tf-PEI and Tf-PEI-PEG polyplex targeting of N2A tumors in A/J mice.

Molecular imaging of the skeleton: quantitative real-time bioluminescence monitoring gene expression in bone repair and development.

Monitoring gene expression in vivo, noninvasively, is a critical issue in effective gene therapy systems. To date, there are no adequate molecular imaging techniques, which quantitatively monitor gene expression in vivo in skeletal development and repair. The aim of this study was to monitor gene expression in skeletal development and repair, using a real-time molecular imaging system, which quantitatively and noninvasively detects bioluminescence in vivo. Our experimental model consisted of transgenic mice harboring the luciferase marker gene under the regulation of the human osteocalcin (hOC) promoter. A new light detection cooled charge coupled device (CCCD) camera was applied to monitor luciferase expression. In vitro, mesenchymal stem cells (MSCs) isolated from bone marrow of transgenic mice exhibited hOC promoter regulation, detected by luciferase expression that correlated with their osteogenic differentiation. During development from 1 week to 1.5 years, transgenic mice exhibited transgene expression in a wide spectrum of skeletal organs, including calvaria, vertebra, tail, and limbs, reaching a peak at 1 week in most of the skeletal organs. In two skeletal repair models, bone fracture and marrow ablation, the noninvasive CCCD system revealed a peak of luciferase expression at 6 days postsurgery. All quantitative, noninvasive, real-time CCCD measurements correlated with a luciferase biochemical assay and luciferase immunohistochemistry, which demonstrated luciferase expression in hypertrophic chondrocytes and trabecular osteoblasts. Our studies show for the first time (1) the CCCD detection system is a reliable quantitative gene detection tool for the skeleton in vivo, (2) expression of luciferase regulated by the hOC promoter is significantly decreased with age in most skeletal sites, and (3) the dynamics of hOC regulation during mice skeletal development and repair in real time, quantitatively and noninvasively.

9] Cytomechanics applications of optical sectioning microscopy

This chapter describes the cytomechanics applications of optical sectioning microscopy. In the past half-century since the purification of actin, the fields of biochemistry and biophysics have produced a vast trove of information on the protein components and regulatory mechanisms of the cytoskeleton. The action or mechanics of the cytoskeleton must be measured and matched to identifiable assemblies. To analyze this cellular machine, an elastic collagen-based model extracellular matrix (ECM) can be utilized as a strain gauge. The light microscope, when suitably automated and interfaced to a digital imaging system, then serves a dual purpose: (1) to image the configuration of the cytoskeleton with molecular specificity through fluorescence labeling and (2) to image the deformations produced by the cell in the surrounding matrix. The basic instrument consists of a fluorescence microscope equipped with a cooled charge-coupled device (CCD) camera system, programmable shutter, grating slider with drive mechanism, and desktop computer for control, image acquisition, and processing. The cooled CCD camera provides the high linearity, dynamic range, geometric precision, and low noise needed for digital processing.