scholarly journals Optimized analysis for sensitive detection and analysis of single proteins via interferometric scattering microscopy

2021 ◽  
Author(s):  
Houman Mirzaalian Dastjerdi ◽  
Mahyar Dahmardeh ◽  
André Gemeinhardt ◽  
Reza Gholami Mahmoodabadi ◽  
Harald Köstler ◽  
...  
Keyword(s):  
Rayleigh Scattering ◽  
Signal To Noise Ratio ◽  
Experimental Setup ◽  
Label Free ◽  
Signal To Noise ◽  
Label Free Detection ◽  
Mass Size ◽  
Technical Issues ◽  
Quantitative Results ◽  
Interferometric Detection

It has been shown that interferometric detection of Rayleigh scattering (iSCAT) can reach an exquisite sensitivity for label-free detection of nano-matter, down to single proteins. The sensitivity of iSCAT detection is intrinsically limited by shot noise, which can be indefinitely improved by employing higher illumination power or longer integration times. In practice, however, a large speckle-like background and technical issues in the experimental setup limit the attainable signal-to-noise ratio. Strategies and algorithms in data analysis are, thus, crucial for extracting quantitative results from weak signals, e.g. regarding the mass (size) of the detected nano-objects or their positions. In this article, we elaborate on some algorithms for processing iSCAT data and identify some key technical as well as conceptual issues that have to be considered when recording and interpreting the data. The discussed methods and analyses are made available in the extensive python-based platform, PiSCAT.

scholarly journals Improvement of SAXS Lab Equipment by Using Scatterless Apertures

2014 ◽  
Vol 70 (a1) ◽  
pp. C1330-C1330
Author(s):  
Joerg Wiesmann ◽  
Andreas Kleine ◽  
Christopher Umland ◽  
André Beerlink ◽  
Juergen Graf ◽  
...  
Keyword(s):  
Signal To Noise Ratio ◽  
Experimental Setup ◽  
High Flux ◽  
Signal To Noise ◽  
Minimum Diameter ◽  
X Ray ◽  
X Ray Scattering ◽  
Wide Range ◽  
Beam Stop ◽  
Synchrotron Beamlines

Parasitic scattering caused by apertures is a well-known problem in X-ray analytics, which forces users and manufacturers to adapt their experimental setup to this unwanted phenomenon. Increased measurement times due to lower photon fluxes, a lower resolution caused by an enlarged beam stop, a larger beam defining pinhole-to-sample distance due to the integration of an antiscatter guard and generally a lower signal-to-noise ratio leads to a loss in data quality. In this presentation we will explain how the lately developed scatterless pinholes called SCATEX overcome the aforementioned problems. SCATEX pinholes are either made of Germanium or of Tantalum and momentarily have a minimum diameter of 30µm. Thus, these novel apertures are applicable to a wide range of different applications and X-ray energies. We will show measurements which were performed either at home-lab small angle X-ray scattering (SAXS) systems such as the NANOSTAR of Bruker AXS or at synchrotron beamlines. At the PTB four-crystal monochromator beamline at BESSY II data was collected for a comparison of conventional pinholes, scatterless Germanium slit systems and SCATEX pinholes. At the Nanofocus Endstation P03 beamline at PETRA III we compared the performance of our SCATEX apertures with conventional Tungsten slit systems under high flux density conditions.

The Noise Analysis of Fluid Systemsin Solid-State Nanopore Sensors

2013 ◽  
Vol 419 ◽  
pp. 517-520 ◽  
Author(s):  
Song Ying ◽  
Lei Wang ◽  
Wen Yuan Zhao
Keyword(s):  
Solid State ◽  
Signal To Noise Ratio ◽  
Noise Analysis ◽  
Power Spectra ◽  
Noise Power ◽  
Label Free ◽  
Signal To Noise ◽  
Fluid Systems ◽  
Noise Ratio ◽  
Nanopore Sensors

The solid-state nanopore sensor offers a versatile platform for the rapid, label-free electrical detection and analysis of single molecules, especially on DNA sequencing. However, the overall signal-to-noise ratio (SNA) is a major challenge in sequencing applications. In our work, two different fluid systems made by metal and plexiglass have been designed to improve the signal to noise ratio of the solid-state nanopore sensor. From the measurements on the noise power spectra with a variety of conditions, it is found that plexiglass fluid system coupled with shielding box produces a good quality of electric signals on nanopore sensors.

scholarly journals Spread spectrum SERS allows label-free detection of attomolar neurotransmitters

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Wonkyoung Lee ◽  
Byoung-Hoon Kang ◽  
Hyunwoo Yang ◽  
Moonseong Park ◽  
Ji Hyun Kwak ◽  
...  
Keyword(s):  
Spread Spectrum ◽  
Signal To Noise Ratio ◽  
Low Cost ◽  
Surface Enhanced Raman Spectroscopy ◽  
Experimental Result ◽  
High Temporal Resolution ◽  
Label Free ◽  
Label Free Detection ◽  
Surface Enhanced Raman ◽  
Sers Detection

AbstractThe quantitative label-free detection of neurotransmitters provides critical clues in understanding neurological functions or disorders. However, the identification of neurotransmitters remains challenging for surface-enhanced Raman spectroscopy (SERS) due to the presence of noise. Here, we report spread spectrum SERS (ss-SERS) detection for the rapid quantification of neurotransmitters at the attomolar level by encoding excited light and decoding SERS signals with peak autocorrelation and near-zero cross-correlation. Compared to conventional SERS measurements, the experimental result of ss-SERS shows an exceptional improvement in the signal-to-noise ratio of more than three orders of magnitude, thus achieving a high temporal resolution of over one hundred times. The ss-SERS measurement further allows the attomolar SERS detection of dopamine, serotonin, acetylcholine, γ-aminobutyric acid, and glutamate without Raman reporters. This approach opens up opportunities not only for investigating the early diagnostics of neurological disorders or highly sensitive biomedical SERS applications but also for developing low-cost spectroscopic biosensing applications.

scholarly journals Highly Multiplexed Label-Free Imaging Sensor for Accurate Quantification of Small-Molecule Binding Kinetics

Proceedings ◽  
2020 ◽  
Vol 60 (1) ◽  
pp. 37
Author(s):  
Elisa Chiodi ◽  
Allison M. Marn ◽  
Matthew T. Geib ◽  
Fulya Ekiz Kanik ◽  
John Rejman ◽  
...  
Keyword(s):  
Small Molecule ◽  
Limit Of Detection ◽  
Signal To Noise Ratio ◽  
Fumonisin B1 ◽  
Biomass Accumulation ◽  
Binding Kinetics ◽  
Label Free ◽  
Common Path ◽  
Label Free Detection ◽  
Imaging Sensor

Investigating the binding kinetics of small molecule analytes to larger ligands, such as proteins and antibodies, is a compelling task for the field of drug and biomarker development, as well as the food industry and agro-biotechnology. Here, we improve the limit of detection of the Interferometric Reflectance Imaging Sensor (IRIS), a label-free, highly multiplexed biosensor, to perform real-time affinity measurement of small molecules binding to immobilized antibodies in a microarray format. As the analytes bind to the surface probes, the biomass accumulation on the surface is quantified by measuring the optical reflectance from the layered Si/SiO2 chip through the solution, in a common-path interferometer configuration. As a proof of concept, label-free detection of biotin molecules binding to immobilized streptavidin probes is performed, achieving 1 pg/mm2 sensitivity through signal averaging in a shot noise limited operation. Furthermore, we apply the optimized sensor to the screening of a 20-multiplexed antibody chip (MW~150 kDa ligands) against Fumonisin B1 (MW = 721.8 Da), one of the most prevalent mycotoxins found in many cereal grains such as corn and wheat. The simultaneously recorded binding curves of the toxin to the multiplexed sensor yield a signal-to-noise ratio of ≈8 when noise reduction methods of spatial and temporal averaging are utilized.

scholarly journals Precision of the Sinefitting-Based Total Harmonic Distortion Estimator

2016 ◽  
Vol 23 (1) ◽  
pp. 37-46 ◽  
Author(s):  
Francisco C. Alegria
Keyword(s):  
Monte Carlo ◽  
Standard Deviation ◽  
Numerical Simulations ◽  
Signal To Noise Ratio ◽  
Harmonic Distortion ◽  
Total Harmonic Distortion ◽  
Experimental Setup ◽  
Square Root ◽  
Signal To Noise ◽  
Monte Carlo Procedure

Abstract An analytical expression for the standard deviation of Total Harmonic Distortion (THD) estimation is derived. It applies to the case where the estimator uses sine fitting. It is shown that, in common circumstances, it is inversely proportional to the actual value of THD, the signal-to-noise ratio and the square root of the number of samples. The proposed expression is validated both with numerical simulations and an experimental setup using a Monte Carlo procedure.

Determination of the Best Emulsion for the Microscopy of Crystals

1981 ◽  
Vol 39 ◽  
pp. 32-33
Author(s):  
David A. Grano ◽  
Kenneth H. Downing
Keyword(s):  
Spatial Frequency ◽  
Information Transfer ◽  
Signal To Noise Ratio ◽  
Experimental Situation ◽  
Photographic Emulsion ◽  
Modulation Transfer ◽  
Signal To Noise ◽  
Frequency Space ◽  
Noise Ratio

The retrieval of high-resolution information from images of biological crystals depends, in part, on the use of the correct photographic emulsion. We have been investigating the information transfer properties of twelve emulsions with a view toward 1) characterizing the emulsions by a few, measurable quantities, and 2) identifying the “best” emulsion of those we have studied for use in any given experimental situation. Because our interests lie in the examination of crystalline specimens, we've chosen to evaluate an emulsion's signal-to-noise ratio (SNR) as a function of spatial frequency and use this as our critereon for determining the best emulsion.The signal-to-noise ratio in frequency space depends on several factors. First, the signal depends on the speed of the emulsion and its modulation transfer function (MTF). By procedures outlined in, MTF's have been found for all the emulsions tested and can be fit by an analytic expression 1/(1+(S/S0)2). Figure 1 shows the experimental data and fitted curve for an emulsion with a better than average MTF. A single parameter, the spatial frequency at which the transfer falls to 50% (S0), characterizes this curve.

Experiments in Bright-Field Imaging with Hollow-Cone Illumination

1981 ◽  
Vol 39 ◽  
pp. 226-227
Author(s):  
W. Kunath ◽  
K. Weiss ◽  
E. Zeitler
Keyword(s):  
Signal To Noise Ratio ◽  
Carbon Support ◽  
Electronic System ◽  
Optic Axis ◽  
Hollow Cone ◽  
Signal To Noise ◽  
Bright Field ◽  
Noise Ratio ◽  
Single Field ◽  
Field Imaging

Bright-field images taken with axial illumination show spurious high contrast patterns which obscure details smaller than 15 ° Hollow-cone illumination (HCI), however, reduces this disturbing granulation by statistical superposition and thus improves the signal-to-noise ratio. In this presentation we report on experiments aimed at selecting the proper amount of tilt and defocus for improvement of the signal-to-noise ratio by means of direct observation of the electron images on a TV monitor.Hollow-cone illumination is implemented in our microscope (single field condenser objective, Cs = .5 mm) by an electronic system which rotates the tilted beam about the optic axis. At low rates of revolution (one turn per second or so) a circular motion of the usual granulation in the image of a carbon support film can be observed on the TV monitor. The size of the granular structures and the radius of their orbits depend on both the conical tilt and defocus.

Correlation averaging of two-dimensional crystals: basic strategy and refinements

1986 ◽  
Vol 44 ◽  
pp. 136-139
Author(s):  
W. Baumeister ◽  
R. Rachel ◽  
R. Guckenberger ◽  
R. Hegerl
Keyword(s):  
Low Dose ◽  
Signal To Noise Ratio ◽  
Real Space ◽  
Fourier Domain ◽  
Two Dimensional ◽  
Signal To Noise ◽  
Unit Cells ◽  
Lateral Displacements ◽  
Established Technique ◽  
Crystalline Array

IntroductionCorrelation averaging (CAV) is meanwhile an established technique in image processing of two-dimensional crystals /1,2/. The basic idea is to detect the real positions of unit cells in a crystalline array by means of correlation functions and to average them by real space superposition of the aligned motifs. The signal-to-noise ratio improves in proportion to the number of motifs included in the average. Unlike filtering in the Fourier domain, CAV corrects for lateral displacements of the unit cells; thus it avoids the loss of resolution entailed by these distortions in the conventional approach. Here we report on some variants of the method, aimed at retrieving a maximum of information from images with very low signal-to-noise ratios (low dose microscopy of unstained or lightly stained specimens) while keeping the procedure economical.

Information capacity and bandwidth limitations in the SEM

1989 ◽  
Vol 47 ◽  
pp. 84-85
Author(s):  
D. C. Joy ◽  
R. D. Bunn
Keyword(s):  
Signal To Noise Ratio ◽  
Critical Dimension ◽  
Information Capacity ◽  
Acquisition Time ◽  
Signal To Noise ◽  
Sem Image ◽  
Probe Size ◽  
Minimum Number ◽  
Noise Ratio ◽  
Signal Channel

The information available from an SEM image is limited both by the inherent signal to noise ratio that characterizes the image and as a result of the transformations that it may undergo as it is passed through the amplifying circuits of the instrument. In applications such as Critical Dimension Metrology it is necessary to be able to quantify these limitations in order to be able to assess the likely precision of any measurement made with the microscope.The information capacity of an SEM signal, defined as the minimum number of bits needed to encode the output signal, depends on the signal to noise ratio of the image - which in turn depends on the probe size and source brightness and acquisition time per pixel - and on the efficiency of the specimen in producing the signal that is being observed. A detailed analysis of the secondary electron case shows that the information capacity C (bits/pixel) of the SEM signal channel could be written as :

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