Analyzing the dynamic range of fluorescent detection in microarray scanner

2006 ◽  
Author(s):  
Wang Liqiang ◽  
Lu Zukang
Keyword(s):  
Dynamic Range ◽  
Fluorescent Detection ◽  
Microarray Scanner

Molecular Beacon-Based Fluorescent Biosensor for Sensitive Detection of Single Nucleotide Polymorphism

2014 ◽  
Vol 651-653 ◽  
pp. 256-259
Author(s):  
Song Bai Zhang ◽  
Meng Tang ◽  
Li Ying Zheng ◽  
Xia Hu ◽  
Guan Gyu Shen ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Fluorescence Intensity ◽  
Molecular Beacon ◽  
Dynamic Range ◽  
High Specificity ◽  
Fluorescent Detection ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Target Dna ◽  
Structure Switching

A sensitive fluorescent biosensing strategy based on structure-switching of molecular beacon for the detection of single nucleotide polymorphism is proposed in this study. In the absence of target DNA, only low background fluorescence can be detected since the formation of hairpin structure induces close proximity of fluorophore and quencher. On the contrary, the fluorescence intensity increases significantly after introduction of target DNA because structure-switching of molecular beacon results in separation of fluorophore and quencher. The change of fluorescence intensity is linear with concentration of target DNA, and a molecular beacon-based sensing system for fluorescent detection of single nucleotide polymorphism is fabricated. The target DNA can be sensitively detected in a linear dynamic range from 17.58nM-1.125μM with a low detection limit of 8 nM. Moreover, good reproducibility and high specificity are achieved.

scholarly journals Quantitative Capillary Electrophoresis for Analysis of Extracellular Vesicles (EVqCE)

Separations ◽  
2021 ◽  
Vol 8 (8) ◽  
pp. 110
Author(s):  
Yuchu Dou ◽  
Lixuan Ren ◽  
Prabir Kumar Kulabhusan ◽  
Emil A. Zaripov ◽  
Maxim V. Berezovski
Keyword(s):  
Capillary Electrophoresis ◽  
Nucleic Acids ◽  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Dynamic Range ◽  
Fluorescent Detection ◽  
Average Mass ◽  
Promising Tool ◽  
Aggressive Cancer ◽  
Capillary Zone

Extracellular Vesicles (EVs) gained significant interest within the last decade as a new source of biomarkers for the early detection of diseases and a promising tool for therapeutic applications. In this work, we present Extracellular Vesicles Quantitative Capillary Electrophoresis (EVqCE) to measure an average mass of RNA in EVs, determine EV concentrations and the degree of EV degradation after sample handling. We used EVqCE to analyze EVs isolated from conditioned media of three cancer cell lines. EVqCE employs capillary zone electrophoresis with laser-induced fluorescent detection to separate intact EVs from free nucleic acids. After lysis of EVs with a detergent, the encapsulated nucleic acids are released. Therefore, the initial concentration of intact EVs is calculated based on a nucleic acid peak gain. EVqCE works in a dynamic range of EV concentrations from 108 to 1010 particles/mL. The quantification process can be completed in less than one hour and requires minimum optimization. Furthermore, the average mass of RNA was found to be in the range of 200–400 ag per particle, noting that more aggressive cancer cells have less RNA in EVs (200 ag per particle) than non-aggressive cancer cells (350 ag per particle). EVqCE works well for the degradation analysis of EVs. Sonication for 10 min at 40 kHz caused 85% degradation of EVs, 10 freeze-thaw cycles (from −80 °C to 22 °C) produced 40%, 14-day storage at 4 °C made 32%, and vortexing for 5 min caused 5% degradation. Presently, EVqCE cannot separate and distinguish individual EV populations (exosomes, microvesicles, apoptotic bodies) from each other. Still, it is tolerant to the presence of non-EV particles, protein-lipid complexes, and protein aggregates.

scholarly journals A Novel Graphene Oxide-Based Aptasensor for Amplified Fluorescent Detection of Aflatoxin M1 in Milk Powder

Sensors ◽  
2019 ◽  
Vol 19 (18) ◽  
pp. 3840 ◽  
Author(s):  
Xiaodong Guo ◽  
Fang Wen ◽  
Qinqin Qiao ◽  
Nan Zheng ◽  
Matthew Saive ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Milk Powder ◽  
High Sensitivity ◽  
High Specificity ◽  
Fluorescent Detection ◽  
Aflatoxin M1 ◽  
Powder Samples ◽  
Infant Milk Powder

In this paper, a rapid and sensitive fluorescent aptasensor for the detection of aflatoxin M1 (AFM1) in milk powder was developed. Graphene oxide (GO) was employed to quench the fluorescence of a carboxyfluorescein-labelled aptamer and protect the aptamer from nuclease cleavage. Upon the addition of AFM1, the formation of an AFM1/aptamer complex resulted in the aptamer detaching from the surface of GO, followed by the aptamer cleavage by DNase I and the release of the target AFM1 for a new cycle, which led to great signal amplification and high sensitivity. Under optimized conditions, the GO-based detection of the aptasensor exhibited a linear response to AFM1 levels in a dynamic range from 0.2 to 10 μg/kg, with a limit of detection (LOD) of 0.05 μg/kg. Moreover, the developed aptasensor showed a high specificity towards AFM1 without interference from other mycotoxins. In addition, the technique was successfully applied for the detection of AFM1 in infant milk powder samples. The aptasensor proposed here offers a promising technology for food safety monitoring and can be extended to various targets.

scholarly journals Tumor Biomarker In-Solution Quantification, Standard Production, and Multiplex Detection

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Nicole C. Japp ◽  
Joshua J. Souchek ◽  
Aaron R. Sasson ◽  
Michael A. Hollingsworth ◽  
Surinder K. Batra ◽  
...  
Keyword(s):  
Dynamic Range ◽  
Fluorescent Detection ◽  
Tumor Biomarker ◽  
Serum Samples ◽  
Liquid Biopsies ◽  
Noninvasive Analysis ◽  
Biomarker Analysis ◽  
Concentration Changes ◽  
Paramagnetic Beads ◽  
New Biomarkers

The diagnosis and monitoring of cancer have been facilitated by discovering tumor “biomarkers” and methods to detect their presence. Yet, for certain cancers, we still lack sensitive and specific biomarkers or the means to quantify subtle concentration changes successfully. The identification of new biomarkers of disease and improving the sensitivity of detection will remain key to changing clinical outcomes. Patient liquid biopsies (serum and plasma) are the most easily obtained sources for noninvasive analysis of proteins that tumor cells release directly and via extracellular microvesicles and tumor shedding. Therefore, an emphasis on creating reliable assays using serum/plasma and “direct, in-solution” ELISA approaches has built an industry centered on patient protein biomarker analysis. A need for improved dynamic range and automation has resulted in the application of ELISA principles to paramagnetic beads with chemiluminescent or fluorescent detection. In the clinical testing lab, chemiluminescent paramagnetic assays are run on automated machines that test a single analyte, minimize technical variation, and are not limited by serum sample volumes. This differs slightly from the R&D setting, where serum samples are often limiting; therefore, multiplexing antibodies to test multiple biomarkers in low serum volumes may be preferred. This review summarizes the development of historical biomarker “standards”, paramagnetic particle assay principles, chemiluminescent or fluorescent biomarker detection advancements, and multiplexing for sensitive detection of novel serum biomarkers.

Slow-Scan CCD Observation of Backscattering Patterns in SEM

1992 ◽  
Vol 50 (2) ◽  
pp. 1308-1309
Author(s):  
F. Ouyang ◽  
D. A. Ray ◽  
O. L. Krivanek
Keyword(s):  
Electron Beam ◽  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Lens System ◽  
Wide Dynamic Range ◽  
Peltier Cooler ◽  
Diffraction Patterns ◽  
Scanning Electron

Electron backscattering Kikuchi diffraction patterns (BKDP) reveal useful information about the structure and orientation of crystals under study. With the well focused electron beam in a scanning electron microscope (SEM), one can use BKDP as a microanalysis tool. BKDPs have been recorded in SEMs using a phosphor screen coupled to an intensified TV camera through a lens system, and by photographic negatives. With the development of fiber-optically coupled slow scan CCD (SSC) cameras for electron beam imaging, one can take advantage of their high sensitivity and wide dynamic range for observing BKDP in SEM.We have used the Gatan 690 SSC camera to observe backscattering patterns in a JEOL JSM-840A SEM. The CCD sensor has an active area of 13.25 mm × 8.83 mm and 576 × 384 pixels. The camera head, which consists of a single crystal YAG scintillator fiber optically coupled to the CCD chip, is located inside the SEM specimen chamber. The whole camera head is cooled to about -30°C by a Peltier cooler, which permits long integration times (up to 100 seconds).

Quantitative energy-filtered electron diffraction

1994 ◽  
Vol 52 ◽  
pp. 992-993
Author(s):  
R. Vincent
Keyword(s):  
Electron Diffraction ◽  
Inelastic Scattering ◽  
Dynamic Range ◽  
Electron Optics ◽  
Surface Layers ◽  
High Brightness ◽  
Inclined Surfaces ◽  
Perfect Symmetry ◽  
Amorphous Surface ◽  
The Ideal

Microanalysis and diffraction on a sub-nanometre scale have become practical in modern TEMs due to the high brightness of field emission sources combined with the short mean free paths associated with both elastic and inelastic scattering of incident electrons by the specimen. However, development of electron diffraction as a quantitative discipline has been limited by the absence of any generalised theory for dynamical inelastic scattering. These problems have been simplified by recent innovations, principally the introduction of spectrometers such as the Gatan imaging filter (GIF) and the Zeiss omega filter, which remove the inelastic electrons, combined with annual improvements in the speed of computer workstations and the availability of solid-state detectors with high resolution, sensitivity and dynamic range.Comparison of experimental data with dynamical calculations imposes stringent requirements on the specimen and the electron optics, even when the inelastic component has been removed. For example, no experimental CBED pattern ever has perfect symmetry, departures from the ideal being attributable to residual strain, thickness averaging, inclined surfaces, incomplete cells and amorphous surface layers.

Direct measurement of electron-diffraction-pattern intensities using an energy loss spectrometer

1989 ◽  
Vol 47 ◽  
pp. 668-669
Author(s):  
A. G. Jackson ◽  
M. Rowe
Keyword(s):  
Thermal Effects ◽  
Dynamic Range ◽  
Scattering Amplitude ◽  
Dynamical Theory ◽  
Site Symmetry ◽  
Proof Of Concept ◽  
Parallel Acquisition ◽  
Kinematic Approximation ◽  
Speed Up ◽  
Structure Calculations

Diffraction intensities from intermetallic compounds are, in the kinematic approximation, proportional to the scattering amplitude from the element doing the scattering. More detailed calculations have shown that site symmetry and occupation by various atom species also affects the intensity in a diffracted beam. [1] Hence, by measuring the intensities of beams, or their ratios, the occupancy can be estimated. Measurement of the intensity values also allows structure calculations to be made to determine the spatial distribution of the potentials doing the scattering. Thermal effects are also present as a background contribution. Inelastic effects such as loss or absorption/excitation complicate the intensity behavior, and dynamical theory is required to estimate the intensity value.The dynamic range of currents in diffracted beams can be 104or 105:1. Hence, detection of such information requires a means for collecting the intensity over a signal-to-noise range beyond that obtainable with a single film plate, which has a S/N of about 103:1. Although such a collection system is not available currently, a simple system consisting of instrumentation on an existing STEM can be used as a proof of concept which has a S/N of about 255:1, limited by the 8 bit pixel attributes used in the electronics. Use of 24 bit pixel attributes would easily allowthe desired noise range to be attained in the processing instrumentation. The S/N of the scintillator used by the photoelectron sensor is about 106 to 1, well beyond the S/N goal. The trade-off that must be made is the time for acquiring the signal, since the pattern can be obtained in seconds using film plates, compared to 10 to 20 minutes for a pattern to be acquired using the digital scan. Parallel acquisition would, of course, speed up this process immensely.

Development of imaging plate for a TEM and its characteristics

1989 ◽  
Vol 47 ◽  
pp. 100-101
Author(s):  
N. Mori ◽  
T. Oikawa ◽  
Y. Harada ◽  
J. Miyahara ◽  
T. Matsuo
Keyword(s):  
Dynamic Range ◽  
Protective Layer ◽  
High Sensitivity ◽  
Imaging Plate ◽  
Phosphor Layer ◽  
Imaging Device ◽  
X Ray Imaging ◽  
New Type ◽  
Basic Characteristics ◽  
Reading System

The Imaging Plate (IP) is a new type imaging device, which was developed for diagnostic x ray imaging. We have reported that usage of the IP for a TEM has many merits; those are high sensitivity, wide dynamic range, and good linearity. However in the previous report the reading system was prototype drum-type-scanner, and IP was also experimentally made, which phosphor layer was 50μm thick with no protective layer. So special care was needed to handle them, and they were used only to make sure the basic characteristics. In this article we report the result of newly developed reading, printing system and high resolution IP for practical use. We mainly discuss the characteristics of the IP here. (Precise performance concerned with the reader and other system are reported in the other article.)Fig.1 shows the schematic cross section of the IP. The IP consists of three parts; protective layer, phosphor layer and support.

Sign in / Sign up

Export Citation Format

Share Document