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At least 361 records · Page 20Linked to original sources
PubMed2004

A faster way to make GFP-based biosensors: two new transposons for creating multicolored libraries of fluorescent fusion proteins.

BACKGROUND: There are now several ways to generate fluorescent fusion proteins by randomly inserting DNA encoding the Green Fluorescent Protein (GFP) into another protein's coding sequence. These approaches can be used to map regions in a protein that are permissive for GFP insertion or to create novel biosensors. While remarkably useful, the current insertional strategies have two major limitations: (1) they only produce one kind, or color, of fluorescent fusion protein and (2) one half of all GFP insertions within the target coding sequence are in the wrong orientation. RESULTS: We have overcome these limitations by incorporating two different fluorescent proteins coding sequences in a single transposon, either in tandem or antiparallel. Our initial tests targeted two mammalian integral membrane proteins: the voltage sensitive motor, Prestin, and an ER ligand gated Ca2+ channel (IP3R). CONCLUSIONS: These new designs increase the efficiency of random fusion protein generation in one of two ways: (1) by creating two different fusion proteins from each insertion or (2) by being independent of orientation.

Biosensing Techniques↗
PubMed2003

Connections between the ivory tower and the multicolored world: linking abstract theories of social justice to the rough and tumble of affirmative action.

This article seeks to combine the social psychologist's interest in articulating and testing concepts with the public policymaker's interest in the effective implementation of specific policies and programs. The first part of the article applies knowledge about distributive and procedural justice to understanding some of the opposition to affirmative action. The application also reveals some lacunae-specifically concerning rule change-in how social psychologists have looked at procedural justice issues. In the second part of the article, we discuss the problem of rule change and propose a set of conceptualizations about the conditions that govern people's reactions to rule change. We end by reflecting on some changes in our studies of procedural justice.

Humans↗
PubMed2002

Generation of multicolor spatial solitons by stimulated Raman scattering.

We report the experimental observation of Raman spatial solitons in a nonlinear planar waveguide. We show that, under certain conditions, stimulated Raman scattering (SRS) can stabilize the pump beam propagation and give rise to multifrequency spatial solitons. We present the different Kerr media with which we obtained self-focusing and SRS. The observations can be described in terms of Raman gain versus soliton power.

Journal Article↗
PubMed2005

Engineering of multicolor spatial solitons with chirped-period quasi-phase-matching gratings in optical parametric amplification.

We demonstrate the excitation of solitons in a parametric amplifier with enhanced signal content through the use of a chirped-period quasi-phase-matching grating. This technique affords a low soliton threshold at the input end of a parametric amplifier, and the subsequent transformation to a desired soliton that exists at nonzero wave-vector mismatch through the use of a linearly chirped quasi-phase-matching grating. This approach has an advantage over direct excitation of solitons at nonzero wave-vector mismatch in uniform nonlinear materials and holds potential for improving the efficiency and mode quality of high-gain parametric amplifiers.

Journal Article↗
PubMed2002

Multicolor FISH.

Explore the source record for details and available documents.

Chromosome Aberrations↗
PubMed2006

New rapid multicolor PRINS protocol.

In the multiple-color primed in situ labeling (multi-PRINS) technique, using ddNTPs between two PRINS reactions can block the free 3'-end generated in the previous PRINS reaction, thus avoiding the next PRINS reaction, using it as a primer to perform spurious elongation at nondesired sites. However, by omitting the blocking step and taking advantage of the color mixing, we developed a simple and rapid multi-PRINS technique to simultaneously detect three chromosomes in the same cell. With this protocol, one can create a third color using the two most common forms of labeled dUTP (biotin- and digoxigenin-labeled dUTP) and two fluorochromes (fluorescein and rhodamine). The signals at the centromeres of three different chromosomes displayed perfect yellow, red, and green colors, respectively. The entire procedure could be completed in less than 90 min because the blocking step was omitted. This protocol is practical and efficient for multi-PRINS so that even more than three chromosome targets could be detected in the same cell.

Chromosomes, Human↗
PubMed2006

Simultaneous multicolor fluorescence cross-correlation spectroscopy to detect higher order molecular interactions using single wavelength laser excitation.

Fluorescence cross-correlation spectroscopy is a powerful method for the study of molecular interactions and dynamics in solution and even in living cells. Usually, in the optical setup, either two laser beams have to be superimposed in their respective confocal volumes or two-photon excitation is used for a dual-color detection system. It has been shown recently that fluorescence cross correlation can be achieved with spectrally similar fluorophores using single wavelength excitation fluorescence cross-correlation spectroscopy (SW-FCCS). In this study, we show that SW-FCCS allows the simultaneous excitation of up to three fluorophores in which the cross correlation of their fluctuation signals is detected separately in three detection channels. The experimental and theoretical model to describe triple pairwise cross correlations incorporating cross talk and possible changes in emission characteristics such as quenching upon binding are outlined. The effectiveness of SW-FCCS to detect binding of three interacting partners is experimentally verified with a standard ligand-receptor model, biotin-streptavidin, where differently labeled biotin ligands and their binding to a third-color labeled streptavidin are studied. The cross-correlation amplitudes and their changes with stoichiometric binding are analyzed and the upper limits of dissociation constants are determined. Performed with appropriate negative controls, SW-FCCS can determine interaction patterns between ligands and receptors.

Biotin↗
PubMed2006

Multicolor quantum dots for molecular diagnostics of cancer.

In the pursuit of sensitive and quantitative methods to detect and diagnose cancer, nanotechnology has been identified as a field of great promise. Semiconductor quantum dots are nanoparticles with intense, stable fluorescence, and could enable the detection of tens to hundreds of cancer biomarkers in blood assays, on cancer tissue biopsies, or as contrast agents for medical imaging. With the emergence of gene and protein profiling and microarray technology, high-throughput screening of biomarkers has generated databases of genomic and expression data for certain cancer types, and has identified new cancer-specific markers. Quantum dots have the potential to expand this in vitro analysis, and extend it to cellular, tissue and whole-body multiplexed cancer biomarker imaging.

Animals↗
PubMed2000

Optical aberrations and objective choice in multicolor confocal microscopy.

Refinements in design have simplified confocal microscopy to the extent that it has become a standard research tool in cell biology. However, as confocal microscopes have become more powerful, they have also become more demanding of their optical components. In fact, optical aberrations that cause subtle defects in image quality in wide-field microscopy can have devastating effects in confocal microscopy. Unfortunately, the exacting optical requirements of confocal microscopy are often hidden by the optical system that guarantees a sharp image, even when the microscope is performing poorly. Optics manufacturers provide a wide range of microscope objectives, each designed for specific applications. This report demonstrates how the trade-offs involved in objective design can affect confocal microscopy.

Animals↗
PubMed2003

Body window-enabled in vivo multicolor imaging of transplanted mouse islets expressing an insulin-Timer fusion protein.

Type 1 diabetes results from the selective destruction of insulin-producing beta cells in the islets of Langerhans, and autoimmune T cells are thought to be the mediators of this destruction. T cells are also responsible for allorejection once the islets are transplanted into a patient to reduce the negative consequences of a lack of insulin. To better understand these processes, we have developed a transgenic mouse expressing proinsulin II tagged with a live-cell fluorescent reporter protein, Timer. Timer protein is unique because it changes color from green to red in the first 24 h after synthesis. With this marker, insulin synthesis can be carefully monitored through fluorescent changes over time. To complement this new biotechnological research tool, we designed a body window to allow for in vivo imaging over time of the islets transplanted under the kidney capsule. The window device, which is sutured to replace the underlying skin and body wall over the site of islet transplantation, may be used to simultaneously observe beta cells and T cells that have been labeled with a fluorochrome distinguishable from Timer. The imaging of both insulin-producing cells and T cells may be carried out repeatedly for a week or more with no need for repeated surgery, while preserving the life of the studied animal.

Animals↗
PubMed2005

Multicolor luciferase assay system: one-step monitoring of multiple gene expressions with a single substrate.

Reporter assays that use luciferase are widely employed for monitoring cellular events associated with gene expression. In general, firefly luciferase and Renilla luciferase are used for monitoring single gene expression. However, the expression of more than one gene cannot be monitored simultaneously by this system because one of the two reporting luciferases must be used as an internal control. We have developed a novel reporter assay system in which three luciferases that emit green, orange, and red light with a single substrate are used as reporter genes. The activities of the luciferases can be measured simultaneously and quantitatively with optical filters. This system enables us to simply and rapidly monitor multiple gene expressions in a one-step reaction.

Animals↗