CytometryML binary data standards

Leif, Robert C.

doi:10.1117/12.589960

Cited by 13 publications

(6 citation statements)

References 9 publications

(7 reference statements)

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“…Under these conditions localized luminescence could be observed, proving that the light-emitting Eu 31 ion of the EuMac had not been exchanged by the non-emitting Gd 31 or Y 31 ions. Fixed apoptotic (65) and S phase (6,66) cells, labeled with the EuMac conjugate of anti-5BrdU, could be imaged with a standard epifluorescence microscope under illumination at 365 nm with a standard Hg-Xe 100 watt arc lamp. Thus, effective imaging could be obtained without the previously needed expensive and complicated timegated or laser-based systems, or without heavy labeling of the anti-5BrdU.…”

Section: Increasing the Luminescence Of Lanthanide Complexes By Energmentioning

confidence: 99%

Increasing the luminescence of lanthanide complexes

et al. 2006

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This review compares the chemical and physical properties of lanthanide ion complexes and of other narrow-emitting species that can be used as labels for cytometry. A series of luminescent lanthanide ion macrocyclic complexes, Quantum Dyes Ò , which do not release or exchange their central lanthanide ion, do accept energy transfer from ligands, and are capable of covalent binding to macromolecules, including proteins and nucleic acids, is described and their properties are discussed.Two methods are described for increasing the luminescence intensity of lanthanide ion complexes, which intrinsically is not as high as that of standard fluorophores or quantum dots. One method consists of adding a complex of a second lanthanide ion in a micellar solution (columinescence); the other method produces dry preparations by evaporation of a homogeneous solution containing an added complex of a second lanthanide ion or an excess of an unbound antenna ligand. Both methods involve the Resonance Energy Transfer Enhanced Luminescence, RETEL, effect as the mechanism for the luminescence enhancement. q 2006 International Society for Analytical Cytology Key terms: Quantum Dye; RETEL; lanthanide; macrocycle; luminescence; columinescence; time-delayed; europium; terbium Only one article published in Cytometry, by Seveus et al.(1), has described in detail the use of a lanthanide ion complex as a luminescent label for an antibody, and this article, published in 1992, dealt primarily with the instrumentation for time-gated microscopy. Thus, it is appropriate for a special issue of Cytometry to include a focused review of this class of luminescent labels. Extensive reviews of the clinical and other uses of lanthanide complexes have previously been published by Hemmil€ a and coworkers (2-4).This review presents a comparison of the spectral properties, relative sizes, and essential chemical features of lanthanide complexes and other narrow emitting labels. It also provides a critical description of two related approaches that can be used to overcome the comparatively low molar extinction coefficients (molar absorptivities) of lanthanide ion complexes: 1) the columinescence effect, where the luminescence of a lanthanide complex is increased in a micellar solution by energy transfer from a complex of a non-emitting lanthanide to a complex of an emitting lanthanide, and 2) the Resonance Energy Transfer Enhanced Luminescence, RETEL, effect (5) where the energy transfer occurs in the solid state.A companion Technical Note (6) describes the experimental aspects of the RETEL effect (5), which resulted in a major increase of the luminescence intensity of a specific type of lanthanide macrocycles, the Quantum Dyes Ò . This increase in luminescence facilitates the use of the Quantum Dyes as labels, either with fluorescent microscopes conventionally illuminated by a Mercury-Xenon (Hg-Xe) arc or-when it is helpful to eliminate contamination from the emissions of conventional fluorophoreswith new cost-effective time-gating instrumentation.The emissions...

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Section: Increasing the Luminescence Of Lanthanide Complexes By Energmentioning

confidence: 99%

Increasing the luminescence of lanthanide complexes

et al. 2006

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“…For the same reason, Figure 3 is presented in pseudocolor, which was produced by a simple subtraction algorithm (16). This software technology should help to increase the clinical acceptance of fluorescence-luminescence cytohistochemistry by clinical professionals.…”

Section: Discussionmentioning

confidence: 99%

“…Both the blue and red images were obtained with the 60 3 oil immersion lens, and were binned to 680 3 518 pixels. Special software (16) written in the Ada programming language generated pseudo-color images of the tissue culture cells labeled with EuMac-anti-5BrdU and of those stained with DAPI. The color image of Figure 3 was finally obtained by subtracting the red (europium) image from the blue and green image planes, and the blue (DAPI) image from the green and red image planes (16).…”

Section: Methodsmentioning

confidence: 99%

Increasing lanthanide luminescence by use of the RETEL effect

et al. 2006

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Background: Luminescent lanthanide complexes produce emissions with the narrowest-known width at half maximum; however, their significant use in cytometry required an increase in luminescence intensity. The companion review, Leif et al., Cytometry 2006;69A:767-778, described a new technique for the enhancement of lanthanide luminescence, the Resonance Energy Transfer Enhanced Luminescence (RETEL) effect, which increases luminescence and is compatible with standard slide microscopy. Methods: The luminescence of the europium ion macrocyclic complex, EuMac, was increased by employing the RETEL effect. After adding the nonluminescent gadolinium ion complex of the thenoyltrifluoroacetonate (TTFA) ligand or the sodium salt of TTFA in ethanol solution, the EuMac-labeled sample was allowed to dry. Both a conventional arc lamp and a time-gated UV LED served as light sources for microscopic imaging. The emission intensity was measured with a CCD camera. Multiple time-gated images were summed with special software to permit analysis and effective presentation of the final image.Results: With the RETEL effect, the luminescence of the EuMac-streptavidin conjugate increased at least six-fold upon drying. Nuclei of apoptotic cells were stained with DAPI and tailed with 5BrdUrd to which a EuMac-anti5BrdU conjugate was subsequently attached. Time-gated images showed the long-lived EuMac luminescence but did not show the short-lived DAPI fluorescence. Imaging of DNA-synthesizing cells with an arc lamp showed that both S phase and apoptotic cells were labeled, and that their labeling patterns were different. The images of the luminescent EuMac and fluorescent DAPI were combined to produce a color image on a white background. This combination of simple chemistry, instrumentation, and presentation should make possible the inexpensive use of the lanthanide macrocycles, Quantum Dyes Ò , as molecular diagnostics for cytological and histopathological microscopic imaging. q 2006 International Society for Analytical Cytology Key terms: quantum dye; columinescence; RETEL; lanthanide; macrocycle; luminescence; time-delayed; europium; terbium Lanthanide ion complexes have found a variety of significant applications in laboratory diagnostics (1-3); however, the weak luminescence of these complexes, which is due to their comparatively low molar extinction coefficients (molar absorptivities), was a major impediment to their use in cytometry (4). The problem of increasing the emission intensity of lanthanide ion complexes has been solved by the use of the resonance energy transfer enhanced luminescence (RETEL) effect, formerly called FRE-TEL (5), combined with measurement in the dry state. In the RETEL effect, as described by Leif, Vallarino, and coworkers in the companion Review article published in this issue of Cytometry (6), light energy originally absorbed by a nonluminescent photon acceptor (a complex or an unbound ligand) is transferred to the central lanthanide ion of a macrocyclic complex, Quantum Dye Ò , thus enhancing its emission int...

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“…A key part of both the Ada and XML software was a few helper Ada packages (classes) and XML schemas, which included the same descriptions of common data-types [20,21], such as numeric types (num_types.xsd), strings (strings.xsd), and files (files.xsd). One important schema is the equivalent of the Dublin Core [22] that has been tailored for software (about.xsd [21]).…”

Section: Schema Development Processmentioning

confidence: 99%

Toward the integration of cytomics and medicine

Leif

2009

Journal of Biophotonics

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The integration of cytomics research and healthcare informatics will facilitate technology transfer and reduce medical costs. The CytometryML prototype of the Advanced Cytometry Standard (ACS) has the benefits of including microscopic image and flow list-mode data, being based on XML and thus is compatible with existing medical and scientific informatics standards, such as HL7, and employing a design based upon the Digital Imaging and Communications in Medicine (DICOM) standard. The reuse of the well tested DICOM model resulted in a great decrease in the design and documentation effort and increased probability of reliability. Schemas for flow cytometers and microscopes have been created. XML schemas for two related types of container (ZIP) files have been specified for a set of measurements. The series and instance containers respectively include the metadata that is constant and the metadata that is specific to an individual or small closely related group of measurements.

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CytometryML binary data standards

Cited by 13 publications

References 9 publications

Increasing the luminescence of lanthanide complexes

Increasing the luminescence of lanthanide complexes

Increasing lanthanide luminescence by use of the RETEL effect

Toward the integration of cytomics and medicine

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